CH515249A - 2 5-and 3 5-dinucleoside phosphates - Google Patents

Abstract

(A) 2',5'- and 3',5'-Dinucleoside phosphates X1 = H, alpha-hydroxy or beta-hydroxy, when oly one of the furanose rings in the same cpd. is a ribofuranose. The combination X2 and X3 = H, alpha-OH; H-beta OH; alpha-OH, beta-OH; beta-OH, alpha-OH; or beta-OH, beta-OH. Y1 and Y2 = cytosin-1-yl, uracil-1-yl, thymin-1-yl, adenin-9-yl, guanin-9-yl, 6-mercaptopurin-9-yl, uracil-3-yl, 5-fluoro-, 5-chloro-, 5-bromo-, 5-iodo- or 5-trifluoromethyl-uracil-1-yl, hypoxanthin-9-yl, xanthin-9-yl, 5-methylcytosin-1-yl or 3-methylcytosin-1-yl. (B) Pharmaceutically acceptable acid addition salts of (I) and (II). The new cpds. have considerable cytotoxic activity in vitro esp. against KB-tumour cells and virus, partic. Herpes-, COe- and vaccine-virus.

Description

  

  
 



  Process for the preparation of 2 ', 5'- and 3', 5'-dinucleoside phosphates
The present invention relates to a process for the production of new 2 ', 5'- and 3', 5'-dinucleoside phosphates and their pharmaceutically usable salts.



   The compounds which can be prepared according to the invention have the following formulas
EMI1.1
 in which X1 denotes hydrogen, α-OH or ss-OH, X3 and X5 each in the combinations H, α-OH; H, ss-OH; α-OH, ss-OH, α-OH or ss-OH, ss-OH, and Y1 and Y are cytosin-1-yl, uracyl-1-yl;

  ; Thymin-1-yl, adenin-9-yl, guanin-9-yl, 6-mercaptopurin-9-yl, uracil-3-yl, 5-fluorouracil-l-yl, 5-chloruracil-1-yl, 5- Bromuracil-1-yl, 5-iodouracil-l-yl, 5-trifluoromethyluracil-1-yl, hypoxanthin-9-yl, xanthin-9-yl, 5-methylcytosin-l-yl or 3-methylcytosine-l- yl, represent.



  If X denotes a-OH or f3-OH, mixtures of compounds of the formulas XI and XII can be obtained.



   The process according to the invention is characterized in that a nucleoside phosphate of the formula
EMI2.1
 in which X 'denotes hydrogen, a-O-acyl or ss-O-acyl, the acyl groups being 2 to 12 carbon
EMI2.2
 Atoms contain and Ac 'is an acyl radical having 2 to 12 carbon atoms with a nucleoside of the formula
EMI2.3
 wherein T is triphenylethyl, (p-methoxyphenyl) -diphenylmethyl or bis- (p-methoxyphenyl) -phenylmethyl, X is hydrogen, a-OH or ss-OH and Y 'corresponds to one of the radicals mentioned for Y1 and Y, in which Amino or imino groups are acylated, in the presence of a dialkylcarbodiimide in the 3'-position, if X is hydrogen, or in the 2'- or 3'-position, if X is sc-OH or -OH, but with the proviso

   that the nucleoside and the nucleoside phosphate, which are to be reacted with one another, are used in the combinations arabinofuranoside-arabinofuranoside, ribofuranoside-arabinofuranoside, deoxyribofuranoside-arabinofuranoside or ribofuranoside-deoxyribofuranoside, in which one obtains a dinucleoside of the formula 'and Xa' have the same meaning as X ', and that the intermediate products obtained are then hydrolyzed first with a weak base and then with an aqueous acid under mild conditions.



   The separation of Y1, Y2, Y1, and Y2 has been made to indicate that these substituents, although derived from the same groups Y and Y ', need not necessarily be identical in compounds VII, VIII, IX and X, i.e. Y1 and Y in compound II can be the same (Yl = Y2), but do not have to be.



   The vertical wavy line with substituents at both ends indicates that the substituents are in both the a-position (i.e. below the plane of the ring) or in the p-position (i.e.



  can be arranged above the ring plane).



   The starting compounds for the process according to the invention can be prepared according to the following reaction scheme: Y cytosin-1-yl, uracil-1-yl, thymin-1-yl (or 5-methyluracil-1-yl), adenin-9-yl ( or 6-aminopurin-9-yl), guanin-9-yl (or 2-amino-6-hydroxypurin-9-yl), 6-mercaptopurin-9-yl, uracil-3-yl, 5-fluorouracil- l-yl, 5-chloruracil-l-yl, 5-bromouracil-l-yl, 5-iodouracil-l-yl, 5-trifluoromethyluracil-l-yl. Hypoxanthin-9-yl (or 6-hydroxypurin-9-yl), xanthin-9-yl (or 2,6-dihydroxypurin-9-yl), 5-methylcytosin-l-yl or 3-methylcytosine -l- yl, Y 'the same radicals as indicated for Y, in which acylatable groups, for example Amino groups, also acylated and thereby protected from a reaction with the phosphate esters in other than the desired position.

  Y 'can therefore be:
EMI3.1

In the above formulas: Ac and Ac 'mean acyl radicals with 2 to 12 carbon atoms, preferably anisoyl radicals, T triphenylmethyl, (p-methoxyphenyl) diphenylmethyl or bis (p-methoxyphenyl) phenylmethyl, X hydrogen, -OH or.p- OH, X 'hydrogen, aO-acyl or 5-O-acyl (where the acyl group corresponds to the definition given above), N4-acylcytosin- l-yl, -uracit- l-yl, -thymin- l-yl, N6- Acyladenin-9-yl, N2-acylguanin-9-yl, -6-mercaptopurin-3-yl, -uracil-3 -yl, -5-fluorouracil-1-yl, -5-chloruracil-1 -yl, -5 -bromuracil- l-yl, -5-ioduracil- l-yl, -5-trifluoromethyluracil-l-yl, -hypoxanthin-9-yl, -xanthin-9-yl,

   N4-acyl-5-methylcytosin-1-yl or N4-acyl-3-methylcytosin-1-yl (the acyl group being as defined above).



   The compounds of the formula VIII can be obtained as follows:
EMI4.1

In the formulas, Ac, T, Y ', X and X' have the meaning given. If the sugar content in the compound III molecule is deoxyribose, i. X '= H, and Y' is not acylated, i.e. if Y '= Y, then the compound of the formula VIII corresponds to the compound of the formula II, and the latter can then be used for the condensation.



   In the compounds of the formula XI and XII which can be prepared according to the invention, X1 denotes hydrogen, a-OH or -OH, with the proviso that only one of the furanose rings in the molecule can be ribofuranose; X and X are each in the combinations H, a-OH ; H, ss-OH; α-OH, ss-OH, α-OH or ss-OH, ss-OH, and Y1 and Y2 have the same meanings as given for Y above.

 

   The new compounds can contain, for example, the following acyl groups: acetyl, propionyl, butyryl, valeryl, isovaleryl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, lauroyl, benzoyl, phenylacetyl, phenylpropionyl, p-toluoyl, 0-cyclopentylpropionyl and others.



   The heterocyclic radicals Y arise when a hydrogen atom is removed from the parent compound at the point indicated by the number before the ending -yl. The radicals Y therefore correspond to the following formulas
EMI4.2
 Cytosin-1-yl uracil-1-yl thymin-1-yl (a) (b) (c)
EMI5.1
 Adenin-9-yl guanin-9-yl (d) (e)
EMI5.2


 <tb> <SEP> SH
 <tb> <SEP> S <SEP> I <SEP> H-N <SEP> ¸
 <tb> <SEP> oXX? \ O
 <tb> 6-mercaptopurine <SEP> in-9-yI <SEP> Uraci <SEP> -3-yl
 <tb> <SEP> (f) <SEP> (9)
 <tb>
EMI5.3
 5-fluorouracil-1-yl 5-chloruracil-1-yl 5-bromouracil-1-yl (h) (i) (j)
EMI5.4
 5-iodouracil-1-yl 5-trifluoromethyl hypoxanthin-9-yl (k) uracil-1-yl (m) (1)
EMI5.5
   Xanthin-9-yl 5-methylcytosin-1-yl 3-methylcytosine (n) (o) 1-yl (p)
The above uracil- (b) and substituted

   Uracil residues (c), (g), (h), (i), (j), (k) and (I) are shown in the keto form and not in the tautomeric enol form.



  Groups other than the above can also be represented in the tautomeric form. For example, the cytosine and substituted cytosine residues (a) and (o) shown above in the amino form can also be shown in the tautomeric imino form.



  Many of the new compounds represent a mixture in which the two possible forms exist in equilibrium.



   The sites which can react in the heterocyclic part and in the sugar part of the molecule with phosphoric acid or a phosphorylating agent are usually protected, while at the same time the 2 'or 3' positions are available for reaction with the phosphorylating agent.



   The starting compounds for the process according to the invention can be prepared as follows:
A compound of the formula I is etherified in the 5'-position in the form of the free base or in the form of the salt with a mineral acid such as hydrochloric acid. e.g. with triphenylchloromethane or a methoxy-substituted triphenylchloromethane, the corresponding 1 - (5 '- O - triphenylethyl, 3-D-furanosyl) compound (II) being obtained. The compound II is then acylated with an acylating agent such as acetic anhydride, propionic anhydride or benzoyl chloride on the hydroxyl groups in the 2 'and 3' positions and, if present, on the amino group of the heterocyclic N-ring (cytosine, adenine). so that the corresponding 1 (2 ', 3'-di.O-acyl-5'.O-triphenylmethyl-p- -D-furanosyl) compound (III) is obtained.

  The compound III is subjected to an acid-catalyzed ether cleavage without further purification, the corresponding 1- (2 ', 3'-di-O-acyl-ss-D-furanosyl) compound (IV) being obtained. The compound IV is then assigned a specifi.



  chemical phosphorylating agents. e.g. 2-cyanoethyl phosphate, treated in the presence of a suitable condensing agent such as dicyclohexylcarbodiimide and then with an alkali such as lithium hydroxide, so that the 19-D-furanosyl-5'-phosphate (V) is obtained. If the compound V is now acylated again, the corresponding 1- (2 ', 3'-di-O-acyl-, 13-D-furanosyl) -5'-phosphate of the formula VI is obtained.



   The dinucleoside phosphates of the formulas XI and XII obtainable according to the invention have a strong cytotoxic activity in vitro. and in particular against KB tumor cells and against viruses, in particular of the Herpes, Coe and Vaccinia species. As a result, the compounds can be used to clean the glassware and instruments used to grow tissue cultures for virus and tumor research. They can also be used to wash excised tumor tissue that is to be transplanted into animals in order to prevent the growth of KB tumor cells that could otherwise get into the surrounding tissue or other parts of the body.

  The antibacterial activity of the compounds prepared according to the invention can also be used to produce phagocyte-free fungi and bacterial cultures, in particular phagocyte-free Streptomyces cultures. In particular, the compounds of formula XII can also be used to treat herpes keratitis in virus-infected domestic animals, e.g. Rabbits, to cure.



   The compound of the formula I used as starting material is a known material (cf., for example, Michaelson The Chemistry of Nucleosides and Nucleotides *, Academic Press, London and New York, 1963, catalogs of the Waldhof cellulose factory, Germany 1964, etc.).



  Further possible starting compounds are mentioned in the examples below.



   In the preparation of the starting compounds, a compound of the formula I in the form of the hydrochloride, the hydrobromide or another salt or as the free base in an organic basic solvent can be treated with an etherifying agent. The following can be used as etherifying agents: triphenylchloromethane, triphenylbromomethane, methoxy-substituted triphenylmethyl bromo- or -chloromethane, for example. (p-methoxyphenyl) diphenylchloromethane, bis (p-methoxyphenyl) phenylchlor or bromoethane. As organic bases pyridine, picoline. Lutidines, ethylpyridines and the like are used; pyridine is preferably used.

  The reaction can be carried out at temperatures between 0 and 60 ° C .; preferably one works at room temperature, i. between about 20 and 300C. The reaction usually takes between 6 hours and 10 days at room temperature. According to a preferred embodiment of this production method, the compound of the formula I can be stirred in pyridine solution together with the approximately equivalent amount of triphenylchloromethane, triphenylbromoethane or their p-methoxy analogs.



   The acylation of the 1 - (5'-O-triphenylmethyl -, 5 - D - furanosvl) compound II obtained in this way can be carried out with acyl chlorides. Acyl bromides and anhydrides of carboxylic acids with 2 to 12 carbon atoms or anisoyl chloride can be made. To the acyl chlorides and acyl halides, which can be used with particular advantage. include: benzoyl chloride, anisoyl chloride, pethylbenzoyl chloride, p-methylbenzoyl bromide, S-cyclopentylpropionyl chloride. Lauryl chloride.



     Decanoyl chloride, octanoyl bromide and the like. Acid anhydrides which can be used advantageously are, for example: acetic acid, propionic acid .. butyric acid, valeric acid, phenylacetic acid. Phenylpropionic and hexanecarboxylic anhydride, etc. According to a preferred embodiment, the reaction can be carried out in dry pyridine at room temperature between 20 and 30 ° C. with continuous stirring; the reaction time is generally about 4 to 48 hours.

  After this time, the reaction mixture can be worked up in the usual way, for example by adding the pyridine solution to water, decanting off the water and purifying the remaining material by chromatography, extraction, recrystallization or a combination of these methods. The 1 - (2'.3'-di-O-acyl-5'.O-triphenylmethyl-ss-D-furanosyl) compound obtained in this way can then be subjected to the ether cleavage by for example with acetic acid or with an acetic acid containing a hydrogen halide, e.g. Contains hydrogen chloride, hydrogen bromide or hydrogen iodide, treated; the 1- (2'.3'-di.O-acyl - $ - D-fura nosyl) compound is obtained in this way.

 

   The phosphorylation of the protected 1- (2 ', 3'-di-O-acyl-ss-D-arabinofuranosyl) cytosine is usually carried out by the method of G. M. Tenor, J. Am. chem. Soc.



  83, 159 (1959). Anhydrous, hydroxyl-free solvents in which the phosphorylating agent, e.g. the phosphate ester, is soluble. These solvents include, for example, pyridine, picoline, lutidine, etc. Neutral solvents such as dimethyl sulfoxide, tetrahydrofuran, N, N-dimethylacetamide or dioxane can also be used, provided that per mole of phosphorylating agent there is one mole equivalent of the basic solvent, e.g. Pyridine, is added. The bases which can be used for this purpose also include the trialkylamines.



   One uses in. general phosphate esters which are easily produced by the action of strong bases, e.g. Alkali hydroxides. The 2-substituted ethyl dihydrogen nhosnhates of the formula are particularly suitable for the reaction
EMI7.1

In this formula: R is hydrogen or a lower alkyl group; Z is a strongly electronegative substituent such as -C
N: -SO2R "; -C-R";
EMI7.2
   -CF2; -CCI3; -CBs; -C1; -Br; + + -F; -NR "3; -NH3; -JO2; -COOR" '; -NO2 and the like; R "is a lower alkyl or aryl group; R" 'is hydrogen or a lower alkyl or aryl group.



   Preferably, 2-cyanoethyl dihydrogen phosphate is used as the phosphate ester.



   Instead of the aforementioned 2-substituted ethyl dihydrogen phosphates, it is also possible, for example, to use o- and p-substituted phenyl dihydrogen phosphates such as o- and p-carboxyphenyl dihydrogen phosphate, o and p-carbamoylphenyl dihydrogen phosphate and o- and> -cyanophenyl dihydrogen phosphate.



   In the solution containing the p-substituted ethyl dihydrogen phosphate or o- or p-substituted phenyl dihydrogen phosphate, the acyl group-protected arabinofuranosylcytosine can be dissolved, if necessary, especially with heating to about 30 up to 50oC. Once the 1- (2 ', 3' -Di-O-acyl-B-D-furanosyl) compound IV has dissolved, a condensing agent is usually added, e.g.



  an alkyl or aryl substituted carbodiimide, preferably dicyclohexacarbodiimide. In addition to carbodiimides, other condensing agents can also be used, for example p-toluenesulfonyl chloride, methoxyacetylene, ketoneimines, trichloroacetonitrile, substituted cyanamides, d-substituted acetonitriles, alkyl and aryl isocyanates, carboxylic acid chlorides, aralkyl chlorocarbonates and others.



   As already indicated, the reaction can preferably be carried out at temperatures which are somewhat above room temperature, i. at temperatures between 20 and 400C. If necessary, it is also possible at lower temperatures, e.g. at around 50C, to work; Likewise, in particularly stored cases, it is possible to increase the reaction temperature up to about 750 ° C. without undesired side reactions occurring. At temperatures between 20 and 40 ° C. and reasonable concentrations, about 4 to 48 hours are generally required for the reaction. Some conversions are usually completed after an hour; in some cases it may take up to 8 days to implement. The greater the dilution of the reaction mixture, the longer the reaction times are usually required.



   The concentration of the reactants is generally not critical. When using equimolecular amounts of l- (2 ', 3'-di-O-acyl-PD-furanosyl) - compound IV, 2-substituted ethyl phosphate and basic catalyst, an approximately quantitative conversion can be achieved if the necessary to complete the reaction The time available is sufficient. In order to shorten the reaction time, the 2-substituted ethyl dihydrogen phosphate in particular is used in an approximately 3 to 4 molar excess over the 1- (2 ', 3'-di-O-acyl-p-D-furanosyl) compound. Once the reaction is complete, a small amount of water can be added to inactivate the excess phosphorylating agent and condensing agent.

  The solution can then be filtered to remove insoluble material; the insoluble material can be, for example, disubstituted ureas, which are formed by reacting the carbodiimides with water. The filtrate can be used for the next stage of the process, namely the cleavage reaction.



   The cleavage can be carried out with an aqueous alkali metal hydroxide solution. In the preferred embodiment, the solution obtained in the first stage, which is 1- (2 ', 3' -di-O-acyl-pD-furanosyl) -5'-yl-2-cyanoethyl phosphate, is generally used contains, initially concentrated to a small volume, preferably under vacuum. It is usually concentrated to such an extent that the residue is in the form of a viscous mass after cooling, then a base, e.g. Add aqueous lithium, sodium or potassium hydroxide with a normality of 0.4 to 2, so that the pH value of the solution rises to 12 to 13, working mainly at temperatures between -10 and + 200C.



  If the reaction is carried out under more vigorous conditions, i.e.



  higher temperatures or longer periods of time, a compound of the formula V in which the substituent is Y1 instead of Y 'can be obtained. When the reaction is complete, the mixture is usually cooled and filtered. The product V can be prepared from the filtrate in a conventional manner, e.g. by extraction, evaporation, precipitation in the form of insoluble phosphate salts, absorption-desorption on resins, recrystallization, etc.

 

   Depending on the reaction conditions under which the basic hydrolysis is carried out, the acyl groups on amino nitrogen, i. at the N4 of the cytosine or substituted cytosine, N6 of the adenine and N2 of the guanine are retained or removed. At lower temperatures from 0 to 200C and with short reaction times, e.g. 10 to 40 minutes, the acyl group on the N4 atom of the cytosine can be retained. If the alkali solution containing the 2-cyanoethyl phosphate is heated to temperatures between 75 and 100 ° C. or if the treatment is continued for longer periods at lower temperatures, the acyl groups can be removed.



      The IPD-furanosylJ'l-, 8-D-furanosyl-5'-dihydrogen phosphate (V) obtained in this way can then be reacylated in the same way as the compound II, usually in anhydrous pyridine and with anhydrides or halides of carboxylic acids works with 2-12 carbon atoms as acylating agent; the corresponding 5'-phosphate of the 1- (2 ', 3'-di-O-acyl-iJ- -D-furanosyl) compound (VI) is usually obtained.



   The compound VIII can be prepared by either acylating a compound of the formula I to give the compound VII, saponifying the compound VII to give the compound VIIa and etherifying the compound VIIa or by saponifying the compound III.



  The etherification and acylation are usually carried out in the same manner as has been described above for the conversion of I to II and of II to III. If an acylated amino group is present, the saponification can be carried out at low temperature. According to a preferred embodiment of the present invention, the saponification is e.g. in a water-alkanol mixture, preferably methyl or ethyl alcohol, which contains so much base that the pH is above 11. Sodium, potassium or lithium hydroxide or ammonium hydroxide is normally used as the base. The precipitates are generally separated off quickly and freed from excess base by washing with water. The product can be further purified by re-treating with base, washing, extracting and recrystallizing.



   The starting product VI obtained in the manner described above is then condensed by the process according to the invention with a furanoside of the formula VIII (a compound of the formula II can also be used if Y1 and Y1 are identical). The condensation is preferably carried out with equimolecular amounts of the two compounds VI and VIII in anhydrous pyridine and in the presence of dicyclohexylcarbodiimide at room temperature, i.e. about 300C through. Instead of pyridine, alkyl-substituted pyridines such as a- ,: P- or γ-methylpyridine, disubstituted and trisubstituted alkylpyridines, dimethylformamide, diethylformamide, etc. can be used. be used.

  The reaction can generally be carried out at temperatures between 0 and 60 ° C., but it is preferred to work at room temperature, i. between about 20 and 300C. The reaction time can be 4 hours to 10 days. The end product is a 3 ', 5'-dinucleoside phosphate of the formula X if X2 (= X) = H, or a mixture of 2', 5'- and 3 ', 5'-nucleoside phosphates of the formulas IX and X, when X in formula II or VIII is OH. The latter mixture can be worked up in the usual way by removing impurities with a water-immiscible solvent, e.g. Petroleum ether, benzene, aSkelly- solve B (technical hexanes), carbon tetrachloride, methylene chloride or ether, extracted and the remaining aqueous reaction mixture lyophilized.

  The extraction and the lyophilization are usually repeated so often that the aqueous reaction mixture is freed from volatile by-products. The products IX and X can be separated, for example, by chromatography on ion exchange resins, solvent extraction in a Craig apparatus, electrophoresis, etc., and optionally further purified by recrystallization, paper chromatography and high-voltage electrophoresis.



   The esters obtained in this way are then treated with a weak base, preferably with anhydrous ammonical methanol or an aqueous base, in order to hydrolyze the acyl groups, and then treated with aqueous acids in order to cleave the ether bond, in this way the dinucleoside phosphates XI are obtained and XII. During hydrolysis, part of the cytosine can lose the acylamino group, so that a uracilyl nucleoside phosphate is formed.



   The products XI and XII can be isolated and purified in the same way as has been described for the compounds IX and X.



   The following preparations and examples serve to further illustrate the present invention. Preparation 1 9-ss-D-arabinofuranosylguanine
EMI9.1


 <tb> <SEP> NHCOCHs <SEP> C7JOCH
 <tb> <SEP> H
 <tb> <SEP> N <SEP> H <SEP> CH
 <tb> H3COC <SEP> + <SEP> Cl
 <tb> N <SEP> HgCl <SEP> H <SEP> H
 <tb> <SEP> C7H70
 <tb> <SEP> \ <SEP> Xylo '
 <tb> <SEP> N <SEP> H2 <SEP> tCOC3 <SEP> t
 <tb> <SEP> HsCOC <SEP> HN <SEP> H3COCN <SEP> < <SEP> N <SEP> < <SEP> N <SEP> J
 <tb> <SEP> C7H70 <SEP> aH2
 <tb> <SEP> C7H7OCX2 <SEP> 0 <SEP> \ <SEP> C7H7O <SEP> CH2
 <tb> <SEP> MeOd / NH <SEP> 0
 <tb> <SEP> 70 \
 <tb> <SEP> H
 <tb> <SEP> HH \
 <tb> <SEP> C7 <SEP> C7
 <tb> <SEP> NH2 <SEP> OH
 <tb> <SEP> H3COC <SEP> N <SEP> H2N <SEP> S <SEP>;

  ;
 <tb> <SEP> HIGH2 <SEP> \ <SEP> 2 <SEP> to HIGH2
 <tb> <SEP> HIGH2
 <tb> <SEP> 0 <SEP> -CH2CO <SEP> 0
 <tb> <SEP> Y111
 <tb> <SEP> H <SEP> aH
 <tb> <SEP> H
 <tb>
The group CTHTO in the above formulas denotes the benzyloxy group
EMI10.1

With mechanical stirring, a suspension of 5.15 g (11.0 millimoles) of the chloromercuric derivative of 2,6-diacetamido-purine [J. Davoll and S.A. Lowry, J.



  At the. Chem. 73, 1650 (1951)] and 4.0 g of purified diatomaceous earth (Celite) in 325 ml of xylene by azeotropic distillation (50 ml). A solution of 4.39 g (10.0 millimoles) of crude, syrupy 2,3,5-tri-O-benzyl-D-arabinofuranosyl chloride [C.P.J. Glaudemans and H.



  G. Fletcher, Jr., J. Org. Chem. 28, 3004 (1963)] in 50 ml of purified xylene was added to the hot first-mentioned suspension with stirring; the mixture was then refluxed for 3 hours with continued stirring and with exclusion of moisture. The hot mixture was filtered through a bed of celite (diatomaceous earth); the filter bed was washed out with hot xylene. The combined filtrates were concentrated in vacuo to a volume of about 100 ml, and the concentrate was added to an excess of Skellysolve B with stirring. The resulting precipitate was separated off, washed with Skellysolve B (the abbreviation SSB is used below for this solvent) and air-dried. The crude product was stirred into chloroform, the mixture was filtered and the filter was carefully washed with chloroform.

  The combined chloroform filtrates were washed three times with 30% strength aqueous potassium iodide solution and twice with water; the organic layer was separated and dried over anhydrous sodium sulfate. The solvent was then removed in vacuo. The residue was stirred up with methanol; the mixture thus obtained was evaporated to dryness in vacuo.



  In this way, 62% of a foamy solid substance was obtained, which was an anomeric mixture in which 9- (2 ', 3', 5'-tri-O-benzyl-ss-D-arabinofuranosyl) - -2,6- diacetamido-purine was the main component.



   A solution of 2.54 g (4.0 millimoles) of this raw material in 100 ml of methanol, which had been saturated with dry ammonia at 0 C, was turned off at 0 C for about 16 hours. The solution was then evaporated to dryness in vacuo; the acetamide was separated by sublimation under reduced pressure. In this way, 1.93 g (81%) of an amorphous solid substance which essentially consists of the S-anomer of 9- (2 ', 3', 5'-tri-O-benzyl-D-arabino furanosyl) -2-acetamido-6-aminopurine.



   By hydrogenolysis of 2.97 g (5.0 millimoles) of the crude monoacetamido derivative (as before) in the method described by Glaudemans and Fletcher [J. Org. Chem. 28, 3004 (1963)] for tri-O-benzyl-pD-arabinofuranosyladenin the manner described and subsequent crystallization from water gave 1.44 g (89%) 9-ss-D-arabinofuranosyl-2-acetamido- 6-aminopurine.



   A solution of 1.30 g (4.0 millimoles) of the above monoacetate and 3.2 g of sodium nitrite in 10 ml of hot water was cooled to ambient temperature and 3.2 ml of glacial acetic acid were added with mechanical stirring until complete dissolution had occurred. Stirring was continued for about an hour, then about the same amount of water was added and stirring was continued for 16 hours at room temperature. The pH of the solution was adjusted to 4 (with pH paper); then the solution was concentrated to dryness in vacuo. The dry residue was stirred up with hot methanol, the suspension was filtered while hot and the filter was washed with hot methanol. The combined methanolic filtrates (approx. 40 ml) were treated with 460 mg (2.0 milligrammatoms) of sodium; then the solution was refluxed for one hour.

  After neutralizing with acetic acid, the solution was concentrated to a volume of 30.40 ml; the resulting slurry was cooled to 50 ° C. for several hours. The crude product was collected, carefully washed with water and finally recrystallized twice from water in the presence of activated charcoal. In this way, 6.77 mg (60%) of 9-p- -D-arabinofuranosyl-2-guanine were obtained in the form of shiny needles.



   Treatment of 9-ss-D-arabinofuranosylguanine with sodium nitrite and acetic acid gives -arabinofuranosylxanthine. In the same way, treatment of 9-D-arabinofuranosyladenine with sodium nitrite and acetic acid gives 9-ss-D-arabinofuranosylhypoxanthine.



   Various ion exchange resins from Dow Co. are used in the following examples.



  These ion exchange resins are as follows: Dowex 50 X 8 Dowex 50 X 8 is a strongly acidic cation exchange resin which contains ring-shaped sulfonic acid exchange groups which are linked to a styrene polymer lattice which is crosslinked with about 8% divinylbenzene .



   Dowex 50W X 8 Dowex 50W X 8 is a specially cleaned Dowex 50 X 8, the resin has a white color in contrast to the yellow-brown color of the Dowex 50 X 8.



   Dowex I X8 Dowex 1 X 8 is a strongly basic anion exchange resin in which quaternary ammonium exchange groups are linked to a styrene polymer lattice.

 

      Dowex AG I X8 Dowex AG 1 X 8 is a specially purified form of Dowex 1 X 8 supplied by Bio-Rod Laboratories, Richmond, California.



   Example I r- (5-o-tril, eiylmthyl-B-D-erabino cytosine (I)
To a solution of 10 g of 1-ss-D-arabinosyl cytosine hydrochloride in 200 ml of pyridine is added 12 g of triphenylchloromethane. The reaction mixture is then stirred for one week at room temperature (23 ° to 260 ° C.). At the end of this time, the reaction mixture is poured into 3 liters of ice water with stirring, whereupon compound 1 separates out as an oil. If the mixture is left to stand overnight, the oil solidifies; the solid product can be filtered off. After breaking, it is carefully washed with water and air-dried.

  The solid substance thus obtained is triturated with 200 ml of boiling heptane; the mixture is filtered, the insoluble material is collected on a sintered glass funnel. The solid substance is then washed several times with 250 ml portions of boiling heptane each time, then dried and added to one liter of boiling acetone, which contains 1 g of activated carbon (Darco C 60). The hot suspension is then filtered in order to remove the activated carbon again. The filtrate is concentrated on a steam bath to a volume of about 75 ml, which is allowed to cool to room temperature. A crystalline product precipitates out. The crystals are collected on a sintered glass funnel and washed once with 25 ml of acetone which has previously been cooled in ice.



  After this treatment, 13 g of compound 1 with F: 227.5-2280C (with decomposition) are present.



  Analysis for C28H27N305:
Calc .: C 69.26 H 5.61 N 8.86
Found: C 69.09 H 5.67 N 8.93
In the same way, 1 '- [5-D- (p-methoxyphenyl) diphenyl-methyl- or 1 - [5' -O-bis (p-methoxyphenyl) phenylmethyl - D - arabinofuranosylcytosine] can be obtained if Reacts cytosine arabinoside or its hydrochloride in pyridine solution with (p-methoxyphenyl) diphenylchloromethane or bis (p-methoxyphenyl) phenylchloromethane at a temperature between 0 and 600C with continuous stirring.



   Compound 1 can also be prepared using triphenylbromomethane instead of triphenylchloromethane.



   Example 2
1- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) uracil (2)
This compound is prepared by reacting 1- (ss-D-arabinofuranosyl) uracil with according to Example 1
Triphenylchloromethane (hereafter with the abbreviation
TPCM) in pyridine.



   Example 3
1- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) thymine (3)
This compound is prepared by reacting 1- (0-D-arabinofuranosyl) thymine with according to Example 1
TPCM in pyridine.



   Example 4
9- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) adenine (4)
This compound is prepared by reacting according to Example 1 from 9- (p-D-arabinofuranosyl) adenine and
Triphenylbromomethane in pyridine.



   Example S.
9- [5'-O- (p-methoxyphenyl) diphenylmethyl-ss-D -arabinofurnnosyl] adenine (5)
The compound 5 is prepared by reaction according to Example 1 from 9- (p-D-arabinofuranosyl) adenine and (p-methoxyphenyl) diphenylchloromethane in pyridine.



   Example 6
9- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) -6-mercaptopurine (6)
Compound 6 is prepared by reaction according to Example 1 from 9- (ss-D-arabinofuranosyl) -6-mercaptopurine and TPCM in pyridine.



   Example 7
1- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) -5-chloruracil (7)
The compound 7 is prepared by reaction according to Example 1 from 1- (ss-D-arabinofuranosyl) -5-chlorouracil and TPCM in pyridine.



   Example 8
1- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) -5-fluorouracil (8)
The compound 8 is prepared by reaction according to Example 1 from 1- (ss-D-arabinofuranosyl) -5-fluorouracil and TPCM in pyridine.



   Example 9
1- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) -5- trifluoromethyluracil (9)
This compound is produced by reaction according to Example 1 from 1- (ss-D-arabinofuranosyl) -5-trifluoromethyluracil and TPCM in pyridine.



   Example 10
1- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) -5-bromuracil (10)
The compound 10 is produced by reaction according to Example 1. I - D-arabinofuranosyl) -5-bromouracil and TPCM in pyridine.



   Example 11
I- (5'-o-Trphnylmthyl-P-D-wabinofurano -S- ioduracil (11)
The compound 11 is prepared by reaction according to Example 1 from 1- (ss-D-arabinofuranosyl) -5-ioduracil and TPCM in pyridine.



   Example 12
9- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) guanine (12)
This compound is prepared by reaction according to Example 1 from 9- (ss-D-arabinofuranosyl) guanine and TPCM in pyridine.



   Example 13
9- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) hypoxanthine (13)
Compound 13 is produced by reaction according to Example 1 from 9- (ss-D-arabinofuranosyl) hypoxanthine with TPCM in pyridine.



   Example 14
9- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) xanthine (14)
This compound is produced by reaction according to Example 1 from 9- (ss-D-arabinofuranosyl) -xanthine with triphenylbromomethane in pyridine.



   The reactions described in Examples 15 to 30 below are also each carried out in the manner described in Example 1 and using pyridine as the solvent.



   Example 15
1- [5'-O- (p-methoxyphenyl) diphenylmethyl-ss-D-arabino furanosyl] -5-methylcytosine (15)
Compound 15 is obtained from 1- (p-D-arabinofuranosyl) -5-methylcytosine and p- (methoxyphenyl) diphenylchloromethane.



   Example 16
1- (5'-O-triphenylmethyl-ss-D-arabinofuranosyl) -3-methylcytosine (16)
Compound 16 is obtained from 1- (p-D-arabinofuranosyl) -3-methylcytosine and TPCM.



   Example 17
9- (5'-O-triphenylmethyl-ss-D-ribofuranosyl) adenine (17)
This compound is obtained from 9- (ss-D-ribofuranosyl) adenine and triphenyl bromoethane.



   Example 18
1- [5'-O- (p-methoxyphenyl) diphenylmethyl-ss-D-ribo furanosyl) -5-methylcytosine (18)
Compound 18 is obtained by reacting 1- (ss-D-arabinofuranosyl) -5-methylcytosine with (p-methoxyphenyl) diphenylchloromethane.



   Example 19
1- (5'-O-triphenylmethyl-ss-D-ribofuranosyl) -5-trifluoro methyluracil (19)
Compound 19 is obtained by reacting 1- (ss-D-ribofuranosyl) -5-trifluoromethyluracil with TPCM.



   Example 20 1- (5'-O-Tripkerzylngethyl -, - D "ribofuranosyl) cytosine (20)
Compound 20 is obtained from 1- (ss-D-ribofuranosyl) cytosine with TPCM.



   Example 21 1- (5 '-0-Tn'phenylmethyl- -D-ribofuranosyl) -3- -methylcytosine (21)
Compound 21 is obtained by reacting I- (B-D-ribofuranosyl) -3-methylcytosine with TPCM.



   Example 22
1- (5'-O-triphenylmethyl-ss-D-deoxyribofuranosyl) uracil (22)
The compound 22 is obtained by reacting I-P-D-deoxyribofuranosyl) uracil with TPCM.



   Example 23
1 - (5 '-0 -Triphenylm ethyl-p -D-deoxyribofuranosyl) - cytosine (23)
This compound is obtained from 1- (ss-D-deoxyribofuranosyl) cytosine and TPCM.



   Example 24
1- (5'-O-triphenylmethyl-ss-D-deoxyribofuranosyl) adenine (24)
The compound 24 can be obtained by reacting 9- (p-D-deoxyribofuranosyl) adenine with triphenylbromo ethane.



   Example 25
I - [5'-O- (p-methoxyphenyl) diphenylmethyl-p-D-deoxyribofurarzasylJ-5-ioduracil (25)
Compound 25 can be obtained by reacting 1- (ss-D-deoxyribofuranosyl) -5-ioduracil with (p-methoxyphenyl) diphenylchloromethane.



   Example 26
1- (5'-O-triphenylmethyl-ss-D-deoxyribofuranosyl) -5- fluoro uracil (26)
This compound is obtained from 1- (ss-D-deoxyribofuranosyl) -5-fluorouracil and TPCM.



   Example 27
1- (5'-O-triphenylmethyl-ss-D-deoxyribofuranosyl) thymine (27)
This compound is obtained from l- (p-D-deoxyribofuranosyl) thymine and TPCM.



   Example 28
9- (5'-O-triphenylmethyl-ss-D-deoxyribofuranosyl) guanine (28)
This compound is obtained from 9- (ss-deoxyribofuranosyl) guanine and triphenylbromomethane.



   Example 29
9- [5'-O- (p-methoxyphenyl) diphenylmethyl-p-D-deoxyribofuranosyl] xanthine (29)
Compound 29 can be obtained by reacting 9- (ss-D-arabinofuranosyl) xanthine with (p-methoxyphenyl) diphenylchloromethane.

 

   Example 30 -O- (p-Methoxyphenyl) diphenylmethyl-ss-D-deoxy ribof uranosyll hypoxa} 2thin (30)
Compound 30 is produced by reacting 9- (ss-D-deoxyribofuranosyl) hypoxanthine with (p-methoxyphenyl) diphenylchloromethane.



   In the same manner as described in Example 1 and the remaining examples above, other heterocyclic 1- [5'-trityl-, 1- [5 '- (p-methoxyphenyl) diphenylmethyl-, and 1- [5'-bis-p Methoxyphenyl) phenylmethyl-pD-ribofuranose and -deoxyribofuranose] compounds are prepared by adding triphenylchloromethane, triphenylbromomethane, (p-methoxyphenyl) diphenylchlor (or bromo) methane or bis (p-methoxyphenyl) -phenylchlor (or bromine) methane with an N-heterocyclic 1 -Q3-D-Rlbofuranosyl) deoxyribofuranosyl connec tion, so that compounds of the formula II are obtained.

  Compounds of the formula II which can be prepared in this manner include the following [in the following enumeration the abbreviation (TPM-RF) is used for the term (5'-O-triphenylmethyl-ss-D-ribofuranosyl) and for the term (5'-O-Triphenylmethyl-BD-deoxyribofuranosyl) the abbreviation (TPM DRF) used]:

   9- (TPM-RF) guanine, 1 - (TPM-RF) -5 -bromuracil, 1 - (TPM-RF) -5-ioduracil, 9- (TPM-RF) hypoxanthine, 9- (TPM-RF) xanthine , 9- (TPM-DRF) hypoxanthine, 1 (TPM-DRF) thymine, 1- (TPM-DRF) -3-methylcytosine, 1 - (TPM-DRF) -5-methylcytosine, 1 - (TPM-DRF) - 5-trifluoromethyluracil, 1 (TPM-DRF) -5-bromouracil, 1 - [5'-O- (p-methoxyphenyl) diphenylmethyl-pD-deoxyribofuranosyl] -5-chloronracil, 9- (TPM-DRF) -6 -mercaptopurine, 1 - [5'-O-bis (methoxyphenyl) phenylmethyl-D-deoxy ribofuranosyl] -3 -methylcytosine, 1- [5'-O-bis (p-methoxyphenyl) phenylmethyl-ss-D-deoxy ribofuranosyl ) -5-methyluracil et al



   Example 31
N4-Benzoyl-1 - (2,3 '-di-0-benzoyl-p -D-arabino-tgrarrosyl) cytosine
A mixture of 6.2 g of 1- (5'-O-triphenylmethylz -D-arabinofuranosyl) cytosine, 40 ml of dry pyridine and.



  6 ml of benzoyl chloride is stirred at room temperature (24 to 260 ° C.) for about 20 hours. The reaction mixture obtained in this way is poured into 500 ml of cold water and stirred at room temperature for 3 hours. The aqueous part of the mixture is then decanted off, the remaining gummy material is washed twice with water, which is decanted off each time.



  The rubbery material and the solid substances are dissolved in 150 ml of methylene chloride; this solution is successively extracted twice with 50 ml of water each time and once with 50 ml of saturated aqueous sodium chloride solution. The methylene chloride solution is then dried by passing it over 10 g of anhydrous sodium sulfate, which is located in a sintered glass funnel. The desiccant was then washed with 20 ml of methylene chloride. The filtrates were combined. The methylene chloride solution was then evaporated at 400 in vacuo. The residue thus obtained was dissolved in 50 ml of chloroform and treated with 6.7 ml of hydrogen bromide in acetic acid (30% hydrogen bromide) with stirring.



  After 3 minutes, the reaction mixture was concentrated to a volume of 10 ml, in vacuo at 40 ° C. The concentrate was diluted with 10 ml of ordinary chloroform and passed through a chromatography column loaded with 100 g of silica gel (180 ml). The silica gel used was a Brinkman silica for chromatographic purposes; the chloroform used was stabilized with hydrocarbon.



  The column was eluted with three times the column volume (540 ml) of chloroform which had been stabilized with ethanol, using a flow rate of about 3.5 ml per minute. The solvents that ran off during this operation were discarded. The column was then eluted with 1.2 ml of chloroform, which had been stabilized with ethanol and mixed with 3 percent by volume of methanol, at a flow rate of 3.5 ml per minute. The solvents drained off during this operation were collected in 20 ml fractions.

  Each fraction was tested for the presence of triphenyl carbinol or triphenyl ether by placing a drop on a sheet of chromatographic paper (Whatman # 40) and examining the stain for ultraviolet absorption after spraying the paper with 5% aqueous sulfuric acid . On the basis of the results of this chromatographic determination, fractions 25 to 43 were combined, washed with 200 mol of water which contained 0.5 ml of pyridine, dried with anhydrous sodium sulfate and evaporated in vacuo. The residue obtained in this way was recrystallized by first dissolving in ethyl acetate and then adding so much SSB that crystallization began; to terminate and complete the crystallization, the mixture was cooled to 40C.

  3 crystal crops could be obtained, all of which were homogeneous, as could be determined by thin layer chromatography using silica gel and a solvent mixture of 10% methanol and 90% benzene. The amount of the three harvests was 1.45 g, 0.940 g and 0.740 g, for a total of 3.13 g (44 SO), the compound 31 obtained in this way had a melting point of 177.5 to 1780 ° C.



  Analysis for C30H2508:
Calc .: C 64.9 H 4.5 N 7.57
Found: C 63.95 H 4.67 N 7.29
Example 32 N4-Acetyl-1- (2 ', 3'-di-O-acetyl-S'-O-triphenylmethyl-, 3-D-rabinofuranosyl) cytosine (32)
A suspension of 750 mg 1- (TPM-AF) cytosine [in this and in the following examples the abbreviation TPM-AF is used for the expression (5'-O-triphenylmethyl-ss-D-arabinofuranosyl)] in 9 ml pyridine treated with stirring with 3 ml of acetic anhydride until a uniform solution was obtained. Stirring was continued for an additional 2 hours during which time the solution turned into a crystalline mass.

  The crystal mass was added to 90 ml of water, the white crystalline material was filtered off, carefully washed with water and dried, in this way 950 mg of crystals with an F: 249-259.50C were obtained.



  After recrystallization from ethanol, 800 g of colorless rosettes of compound 32 with F: 251-2520C were present.



  Analysis for Cs4H3sO7Ns
Calc .: C 66.76 H 5.44 N 6.87
Found: C 67.04 H 5.47 N 7.00
Example 33
N4- (ss-D-Cyclopentylpropionyl) -1- [2 ', 3'-di-O- (ss-cyclopentylpropionyl) - -D-arabinofuranosyfl (33) (33)
In the manner described in Example 31, 1- (TPM-AF) cytosine was treated with cyclopentylpropionyl chloride in pyridine and then with hydrogen bromide which was dissolved in acetic acid. In this way compound 33 was obtained.



   Example 34
N4-Lauroyl-1- (2 ', 3'-di-O-lauroyl-ss-D-arabino furanosyl) cytosine (34)
Compound 34 is obtained if, in the manner described in Example 31, 1- (TPM-AF) cytosine is reacted with lauroyl chloride in pyridine and then with hydrogen bromide in acetic acid.



   Example 35 N4-Propionyl-1- (2 ', 3'-di-O-propionyl-5'-triphenylmethyl -ss-D-arabinofuranosyl) cytosine (35)
Compound 35 is obtained if 1- (TPAI: -AF) cytosine is reacted with propionic anhydride in pyridine in the manner described in Example 32.



   Example 36
N4-butyryl-9- (2 ', 3'-di-O-butyryl-5' -0-triphenylmethyl- -p -D-arabinofurannsyl) adenine (36)
Compound 36 is obtained when 9- (TPM-AF) adenine is reacted with butyric anhydride in pyridine in the manner described in Example 32.



   Example 37 1- (2 ', 3' -Di-O-phenylacetyl -5 '-O-triphenyZmethyl - -ss-D-arabinofuranosyl) thymine (37)
The compound 37 can be obtained if 1 - (TPM-AF) - thymine is reacted with phenylacetic anhydride in pyridine in the manner described in Example 32.



   Example 38 1- (2 ', 3' -Di-O-hexanoyl-5'-O-triphenylmethyl-p-D- -arabinof uranosyl) uracil (38)
If 1- (TPM-AF) uracil is reacted with hexanecarboxylic anhydride in pyridine in the manner described in Example 32, compound 38 is obtained.



   Example 39 9- (2 ', 3'-Di-O-pherzylpropionyl-5'-O-tr.iphenylmethyl- -p-D-arabinofuranosyl) xarethiri (39)
If 9- (TPM-AF) xanthine is reacted with phenylpropionic anhydride in pyridine in the manner described in Example 32, the compound 39 is obtained.



   Example 40
1- (2 ', 5'-Di-O-benzoyl-ss-D-arabinofuranosyl) -5 -chloruracil (40)
In the manner described in Example 31 was
1- (TPM-AF) -5-chloruracil is reacted with benzoyl chloride, and the resulting product was treated with a solution of hydrogen bromide in acetic acid. In this way compound 40 was obtained.



   Example 41
N6-Benzoyl-9- (2 ', 3'-di-O-benzoyl-ss-D-arabino furanosyl) guanine (41)
9- (TPM-AF) guanine was reacted with benzoyl chloride in the manner described in Example 31, and the reaction product thus obtained was treated with a solution of hydrogen bromide in acetic acid. In this way compound 41 was obtained.



   Example 42
9- (2 ', 3'-di-O-benzoyl-ss-D-arabinofuranosyl) -6 -mercaptopurine (42)
Reaction of 9- (TPM-AF) -6-mercaptopurine with benzoyl chloride in the manner described in Example 31 and further treatment of the product thus formed with a solution of hydrogen bromide in acetic acid gave compound 42.



   Example 43
9- (2 ', 3'-Di-O-acetyl-5'-O-triphenylmethyl-ss-D -arabinofurarrosyl) xargthin (43)
This compound can be obtained when 9- (TPM-AF) uracil is reacted with acetic anhydride in the manner described in Example 32.



   Example 44 1- (2 ', 3' -Di-O-phenylacetyl-5 '-O-triphenylmethyl- -ss-D-arabinofuranosyl) -5-fluorouracil (44)
The compound 44 can be obtained if 1- (TPM-AF) -5-fluorouracil is reacted with phenylacetic anhydride in the manner described in Example 32.



   Example 45
N4-valeryl-9- (2 ', 3'-di-O-valeryl-5'-O-triphenylmethyl -ss-D-arabinofuranosyl) -5-methylcytosine (45)
If 9- (TPM-AF) -5-methylcytosine is reacted with valeric anhydride in the manner described in Example 32, compound 45 is obtained.



   Example 46
1- (2 ', 3'-Di-O-benzoyl-ss-D-ribofuranosyl) uracil (46)
If l- (TPM-AF) uracil is reacted with benzoyl chloride in the manner described in Example 31 and the product thus formed is reacted with a solution of hydrogen bromide in acetic acid, compound 46 is obtained.



   Example 47
N4-Benzoyl-1- (2 ', 3'-di-O-benzoyl-ss-D-ribofuranosyl) cytosine (47)
If l- (TPM-AF) cytosine is reacted with benzoyl chloride in the manner described in Example 31 and the product thus formed is reacted with hydrogen bromide in acetic acid, compound 47 is obtained.



   Example 48
1- (2 ', 3'-Di-O-benzoyl-ss-D-ribofuranosyl) thymine (48)
It is set in the manner described in Example 31
1- (TPM-AF) thymine with benzoyl chloride and the resulting product with a solution of hydrogen bromide in acetic acid, the compound 48 is obtained.



   Example 49
1- (2 ', 3'-Di-O-lauroyl-ss-D-ribofuranosyl) -5-fluorouracil (49)
To obtain compound 49, 1- (TPM-AF) -5-fluorouracil is first reacted with lauroyl chloride in the manner described in Example 31 and the product thus formed is treated with a solution of hydrogen bromide in acetic acid.



   Example 50 N0-decanoyl-9- (2,3 '-di-0-decanoyl-D-nbo-furanosyl) adenine (50)
One sets in the manner described in Example 31
9- (TPM-AF) adenine with decanoyl chloride and treated the resulting product with a solution of
Hydrogen bromide in acetic acid, in this way compound 50 is obtained.



   Example 51
9- (2 ', 3, -Di-O-octanoyl -0 -D-ribofuranosyl) -6- -mercaptopurine (51)
Treated in the manner described in Example 31 9- (TPM-AF) -6-mercaptopurine first with Oc tanoyl chloride and then with hydrogen bromide in acetic acid. In this way connection 51 is obtained.



   Example 52
1- (3'-O-Benzoyl-ss-D-deoxyribofuranosyl) uracil (52)
In the manner described in Example 31, 1- (TPM-DRF) uracil is reacted with benzoyl chloride and the product thus formed is reacted with a solution of hydrogen bromide in acetic acid. Compound 52 is obtained as the end product.



   Example 53
N4-Benzoyl-1- (3'-O-Benzoyl-ss-D-deoxyribofuranosyl) cytosine (53)
In the manner described in Example 31, 1- (TPM-DRF) cytosine is first reacted with benzoyl chloride and the product thus obtained is reacted with hydrogen bromide in acetic acid. In this way connection 53 is obtained.



   Example 54
1- (3'-O-Benzoyl-ss-D-deoxyribofuranosyl) -5-fluorouracil (54)
By reacting 1- (TPM-DRF) -5-fluorouracil first with benzoyl chloride and then with a solution of hydrogen bromide in acetic acid in the manner described in Example 31, compound 54 is obtained.



   Example 55 N-Acetyl-1- (3'-O-acetyl-P-D-deoxyriof -5-methylcytosine (55)
By reacting 1- (TPM-DRF) -5-methylcytosine first with benzoyl chloride and then with hydrogen bromide in acetic acid in the manner described in Example 31, compound 55 is obtained.



   Example 56
9- (3'-O-Benzoyl-ss-D-deoxyribofuranosyl) -6 -mercaptopurine (56)
By reacting 9- (l? PM-DRF) -6-mercaptopurine with benzoyl chloride and the resulting product with hydrogen bromide in acetic acid in the manner described in Example 31, the compound 56 is obtained.



   Example 57
1- (2 ', 3'-Di-O-acetyl-5'-triphenylmethyl-ss-D-ribo furanosyl) uracil (57)
If, as described in Example 32, 1- (TPM -RF) uracil is reacted with acetic anhydride, compound 57 is obtained.



   Example 58 I- (2 ', 3' Di-O-acetyl-5'-triphenylmethyl-fi-D-ribofuranosyl) -5-fluorouracil (58)
It is set in the manner described in Example 32
1- (TPM-RF) -5-fluorouracil with acetic anhydride, the compound 58 is obtained.



   Example 59
9- (2 ', 3'-di-O-propionyl-5'-triphenylmethyl-ss-D-ribo furanosyl) -6-mercaptopurine (59)
By reacting 9- (TPM-RF) -6-mercaptopurine with acetic anhydride in the manner described in Example 32, the compound 59 is obtained.



   In the manner described in Example 31, other acyl compounds of the formula IV can also be prepared by reacting compounds of the formula II with acid anhydrides, acyl chlorides or bromides (with 2 to 12 carbon atoms) or anisic acid and the 5'-ether group with hydrogen halide, in particular Hydrogen bromide or hydrogen iodide splits.

  The following compounds can be obtained, for example: N4-lauroyl-9- (2 ', 2'-di-O-lauroyl-lp-D-arabinofuranosyl) adenine, 9- (2', 3'-di-O-valeryl- ss-D-ribofuranosyl) hypoxanthine, 9- (2 ', 3'-Di-O-hexanoyl-ss-D-ribofuranosyl) xanthine, 9- (2', 3'-Di-O-octanoyl-ss-D- ribofuranosyl) -3-uracil, 9- (2 ', 3 -Di-O-isobutyryl-PD-nbofurano syl) -5-fluorouracil, 1- (2', 3'-Di-O-anisoyl-ss-D- ribofuranosyl) thymine, N4-phenylacetyl-1- (2 ', 3'-di-O-phenylacetyl-ss-D-ribofuranosyl) -3 -methylcytosine, 1- (3'-O-butyryl-ss-D-deoxyribofuranosyl ) -5-ioduracil, 1- (3'-O-Undecanoyl-pD-deoxyribofuranosyl) -5-trifluoromethyluracil, 1- (3'-O-decanoyl-ss-D-deoxyribofuranosyl) -5-bromo uracil, 9- (3'-O-heptanoyl-ss-D-deoxyribofuranosyl) guanine, 9- (3 '-O-nonanoyl-pD-deoxyribofuranosyl) -6-mercaptopurine, 9- (3'-O-octanoyl-ss-D -deoxyribofuranosyl) xanthine and others

 

   Example 60
1- (3'-O-propionyl-5'-triphenylmethyl-ss-D-deoxy ribofuranosyl) uracil (60)
Compound 60 can be obtained if 1- (TPM-DRF) uracil is reacted with propionic anhydride in the manner described in Example 32,
Example 61
1- (3'-O-butyryl-5'-triphenylmethyl-ss-D-deoxy ribofuranosyl) -5-fluorouracil (61)
To obtain compound 61, 1- (TPM-DRF) -5- -fluorouracil is reacted with butyric anhydride in the manner described in Example 32.



   Example 62
N4-acetyl-1- (2 ', 3'-di-O-acetyl-ss-D-arabinofuranosyl) cytosine (62) and 1- (2', 3'-di-O-acetyl-ss-D-arabinofuranosyl ) cytosine (62a)
A suspension of 10 ml of 80% aqueous acetic acid and 1.3 g of the 5'-triphenylmethyl derivative of the compound 62 is heated to reflux for 10 minutes. The suspension is then cooled, freed of precipitated triphenylcarbinol by filtration and in vacuo at a temperature between 30 and 400C evaporated. The residue is taken up with 20 ml of methanol and poured over a column with 200 ml of silica gel. The column is then eluted with thirty 20 ml fractions (methanol 2517o, benzene 75%).



  Fractions 5 to 11 are combined and recrystallized from acetone-SSB; in this way 240 ml of a substance with F: 171-172.50C are obtained. After repeated recrystallization, the pure compound 62 with F: 174.5 to 175.50C is obtained.



  Analysis for C16Hl308N3:
Calc .: C 48.78 H 5.19 N 11.38
Found: C 48.79 H 4.81 N 11.66
Fractions 26-29 contained a small amount of compound 62a.



   Example 63
N4-anisoyl-1- (2 ', 3'-di-O-benzoyl-ss-D-arabino furanosyl) cytosine (63) A. N4-anisoyl-1-ss-D-arabinofuranosylcytosine (63a)
5 g of 1-p-D-arabinofuranosylcytosine and 25 ml of anisoyl chloride were dissolved in 100 ml of pyridine; the solution was stirred at about 250 ° C. for 6 hours. This mixture was then added with 400 ml of 1.5N hydrochloric acid and allowed to stand overnight at room temperature, i. between 22 and 240C. After this time the precipitated solid substance was filtered off, washed, carefully triturated with water and air-dried.



  The dry substance was suspended in a mixture of 275 ml of water and 251 ml of ethanol and heated to 70 ° C. on a steam bath. The mixture was then cooled to 40 ° C. and the pH was adjusted to 8 by adding 1N sodium hydroxide solution. The solid substance was immediately filtered off, washed with water, air-dried, washed again with 300 ml of ether, filtered and air-dried. Yield of crude compound 63a: 16.6 g. The crude product was taken up in 195 ml of pyridine and 65 ml of water and cooled with ice. 350 ml of 1.5N sodium hydroxide were then added to the solution, cooled to the temperature of the ice bath, over the course of half an hour, with vigorous stirring.

  The reaction was terminated by adding 350 ml of Dowex 50 X 84 (fineness: 450-1600 meshes / cm2) pyridinium resin and stirring for 20 minutes (pH 7.0). The insoluble material was then filtered off from the solution; the residue was washed with water. The combined filtrates were evaporated to dryness in vacuo at 50.degree. The residue was triturated three times with 200 ml of ether each time and filtered. The solid substance was then suspended in 300 ml of boiling water and filtered three times. The combined filtrates were concentrated to a small volume under reduced pressure; In this way, 2.0 g of a product with an F: 197-2000 ° C. (with decomposition) were obtained.

  This raw material was recrystallized four times from water and once from ethanol; then the pure compound 63a with F: 200.5-201.50C (with decomposition) was present.



  Analysis for C17H19O7N2;
Calc .: C 54.11 H 5.08 N 11.14
Found: C 54.38 H 4.82 N 11.31 B. N4-anisoyl-1- [5'-O- (p-methoxyphenyl) diphenyl methyl-ss-D-arabinofuranosyl] cytosine (63b)
A solution of 4.8 g of compound 63a in 50 ml of pyridine was treated with (p-methoxyphenyl) diphenylchloromethane. After 9 hours, 10 ml of methanol were added and the pyridine solution was poured into 600 ml of water with stirring. As soon as the gummy substance formed had coagulated, the solution was poured off, the gum was washed several times with water (decanting each time) and then taken up in methylene chloride. The methylene chloride solution was washed twice with water and once with saturated sodium chloride solution.

  After evaporation of the washed and dried over anhydrous sodium sulfate solution in vacuo at 300C, a residue remained, which was dissolved in benzene and passed through a column of silica gel (5.8 × 48 cm). The column was eluted as follows: twenty 100 ml fractions with 2% methanol and 98% benzene, and 40 100 ml fractions with 5% methanol and 95% benzene. Fractions 49-60 gave a solid crystalline substance after trituration with ether, which was separated off and washed with ether. In this way, 4.21 g of the crude compound 63b was obtained.



  C. Compound 63
4 g of compound 63b in 20 ml of dry pyridine were treated with 3 ml of benzoyl chloride. The closed flask containing the reaction mixture was placed at room temperature for 18 hours. The reaction mixture was then poured into ice water and kept at about 250 ° C. for 3 hours. stirred, with a gummy substance being deposited. This crude product was extracted twice with 50 ml of methylene chloride; the extracts were combined, washed five times with water and once with saturated aqueous sodium sulfate solution and evaporated to dryness in vacuo. The residues were codistilled with toluene under reduced pressure to remove residual pyridine. The residue purified in this way was taken up in 50 ml of dioxane and treated with 80% acetic acid.

  Sufficient hydrochloric acid was then added to the solution to make a 0.03N solution. Finally, the reaction mixture was left to stand for 5 hours. The solvents were then distilled off in vacuo at 40 ° C. and the residue was treated with 100 ml of a chloroform-ethanol mixture (1: 1), which was then evaporated again. The residue was taken up again in chloroform and passed through a silica gel column (2.8 cm diameter, 40 cm height and 250 ml column volume). The column was eluted four times with 250 ml fractions of chloroform containing 0.75% ethanol and then with six 250 ml fractions of chloroform containing an additional 3% methanol. Fractions 5 to 8 were combined and absorbed on a silica gel column (2.8 cm X 50 cm).

  This column was then eluted with four times the column volume of ordinary chloroform and subsequently with 2 liters of a 3% strength methanol solution in chloroform, 20 ml fractions being collected at a dropping rate of 5 ml per minute. Fractions 46-54 (220 ml) contained the desired material which was recrystallized from ethyl acetate-SSB; F: 172-1730C; this material represented the pure compound 63.



  Analysis for C21H27N3O9;
Calc .: N 7.18
Found: N 7.23
In the manner described in Example 32, other acylated and 5'-etherified compounds of the formula III can be prepared by treating a compound of the formula II with an acyl halide, an acid anhydride or anisic acid as acylating agent.

  In this way the following compounds can be obtained: 1- (2 ', 3'-Di-O-heptanoyl-TPF-RF) -5-ioduracil; N4-phenylacetyl- 1 - (2 ', 3' -di-O-phenylacetyl-, N4-hexanoyl-1- (2 ', 3'-hexanoyl-, N4-phenylpropionyl-1- (2', 3'-di -O-phenylpropionyl-, N4-butyryl- 1- (2 ', 3'-di-O-butyryl-, N4-valeryl- 1- (2', 3 -di-O-valeryl-, N4-hexanoyl-1 - (2 ', 3'-di-O-hexanoyl-, N4-heptanoyl-1- (2', 3'-di-O-heptanoyl- and N4-octanoyl- 1 - (2 ', 3'-di- O-octanoyl-TPM-AF) cytosine; N4-acetyl-1- [2 ', 3'-di-O-acetyl-5'-O-, N4-acetyl-1- [2', 3'-di- O-acetyl-5'-O-bis- and N4-phenylpropionyl-1- [2 ', 3'-di-O-phenylpropionyl-5'-O - (p-methoxyphenyl) diphenylmethyl-ss-D-arabinofura nosyl] cytosine; N4-valeryl-1- (2 ', 3'-di-O-valeryl-TPM-RF) cytosine; l- (2', 3'-di-O-hexanoyl-TPM-AF) uracil; 9- (2 ', 3'-di-O-heptanoyl-TPM-DRF) xanthine;

   1- (2 ', 3'-di-O-octanoyl-TPM-AF) guanine; N6-acetyl-1 - [2 ', 3' -di-O-acetyl-5 '-O- (p-methoxyphenyl) - diphenylmethyl-ss-D-deoxyribofuranosyl] adenine; N4-acetyl-1- [2 ', 3'-di-O-acetyl-5'-O-bis (p-methoxyphenyl) phenylmethyl-ss-D-deoxyribofuranosyl] -3-methyl cytosine; 1- [2 ', 3'-Di-O-phenylpropionyl-5'-O- (p-methoxyphenyl) diphenylmethyl-ss-D-ribofuranosyl] -5-chlorouracil; N6-valeryl-1- (2 ', 3'-di-O-valeryl-TPM-AF) -3-methyl cytosine; N4-hexanoyl- 1- (2 ', 3'-di-O-hexanoyl-TPM-RF) -5-methylcytosine:

   9- (2 ', 3'-di-O-heptanoyl-TPM-DRF) hypoxanthine; 1 - (2 ', 3' -Di-O-octanoyl-TPM-AE) -5'-trifluoromethyluracil; N2-acetyl-9- [2 ', 3'-di-O-diacetyl-5'-O- (p-methoxyphenyl) -diphenylmethyl-ss-D-ribofuranosyl] guanine; N6-acetyl- [2 ', 3'-di-O-acetyl-5'-O-bis (p-methoxyphenyl) phenylmethyl-ss-D-arabinofuranosyl] adenine; N4-anisoyl-1 - [2 ', 3'-di-O-anisoyl-5'-O- (p-methoxyphenyl) diphenylmethyl-ss-D-deoxyribofuranosyl] -5-methyl cytosine et al.



   Example 64
N4-butyryl-1- (2 ', 3'-di-O-butyryl-ss-D-arabino furanosyl) cytosine (64)
Compound 64 is obtained when N4-butyryl-1- (2 ', 3'-di -O-butyryl-TPM-AF) cytosine is heated in aqueous acetic acid in the manner described in Example 62.



   Example 65 N4-Phenyiacetyl-1 - (2 ', 3' -di-O-phenyCacRtyl-p-D- -arsbinaf urmosyl) cytosirz (65)
Compound 65 is obtained when N4-phenylacetyl-1- (2 ', 3-di-O-phenylacetyl-TPM-AF) cytosine is heated in aqueous acetic acid in the manner described in Example 62.



   Example 66 N4-Hexanoyl-1- (2 ', 3' -di-O-hexanoyl-ss-D -arabinofurarmsyl) cytosine (66)
Compound 66 is obtained when N4-hexanoyl-1- (2 ', 3' -di-O-hexanoyl-TPM-AF) cytosine is heated in aqueous acetic acid in the manner described in Example 62.



   Example 67
N4-phenylpropionyl-1- (2 ', 3'-di-O-phenylpropionyl -ss-D-arabinofuranosyl) cytosine (67)
Compound 67 is obtained when N4-phenylpropionyl-1- (2 ', 3'-di-O-phenylpropionyl-TPM-AF) cytosine is heated in aqueous acetic acid in the manner described in Example 62.



   In the same manner as described in Example 62, other N-acyl-1- (2 ', 3'-di-O-acyl-ss-D-arabinofuranosyl) cytosines can be prepared by adding the corresponding N4 - acyl - 1 - (2 ', 3'-di-O-acyl-5'-O-triphenylmethyl-ss-D-arabinofuranosyl) cytosines heated with aqueous acetic acid. The following connections can be made, for example:

  : N4-valeryl-1- (2 ', 3'-di-O-valerylss-D-arabinofuranosyl) - cytosine' Nê-hexanoyl-1- (2 ', 3'-di-O-hexanoyl-ss-D- arabinofura nosyl) guanine, NG-heptanoyl-1- (2 ', 3'-di-O-heptanoyl-ss-D-arabinofura- nosyl) adenine, N4-octanoyl-1- (2', 3'-di-O -octanoyl-ss-D-arabinofura nosyl) -3-methylcytosine, Nt4-benzoyl-1- (2 ', 3'-di-O-benzoyl-, 8-D-arabinofuranosyl) - cytosine, Ng-anisoyl-9- (2 ', 3' -di-O-anisoyl-B -D-arabinofuranosyl) - adenine,

   9- (2 ', 3'-di-O-lauroyl-ss-D-ribofuranosyl) xanthine, N4-octanoyl-1- (2', 3'-di-O-octanoyl-pD-arabinofuranosyl) -3 -methylcytosine, N2-decanoyl-9- (2 ', 3'-di-O-decanoyl-pD-arabinofura- nosyl) guanine, N4-butyryl-1- (2', 3'-di-O-butyryl-ss -D-arabinofuranosyl) -5-methylcytosine, 9- (2 ', 3'-di-O-phenylpropionyl-pD-ribofuranosylt-6-mercaptopurine, N6-propionyl-1- (2', 3'-di-O -propionyl-ss-D-arabinofura nosyl) -5-methylcytosine, 1 - [2 '3' -Di-O- (P-cyclopentyIpropionyl), BD-rib syl) -5-ioduradl, 1 - (2 ', 3 '-Di-O-anisoyl-ss-D-ribofuranosyl) -5-fluorouracil,

   1 - (3 '-O-valeryl-, 8-D-deoxyribofuranosyl) uracil, 1 - (2', 3'-di-O-benzoyl-sS-D-ribofuranosyl) -5-trifluoromethyluracil, 1 - ( 3'-O-hexanoyl-pD-deoxyribofuranosyi) thymine et al



   Example 68 1 p-D-Arabinofuranosylytosine 5'-phosphate (68)
To a solution of 40 ml of pyridine and 0.325 m 2 of cyanoethyl phosphate was added 2.5 g of N4-acetyl-1- (2 ', 3'-di -O-acetyl-pD-arabinofuranosyl) cytosine, which contains a small amount of 1- ( 2 ', 3'-Di-O-acetyl-, 3-D-aribonfuranosyl) cytosine. A further 20 mol of pyridine containing 5.6 g of dicyclohexylcarbodiimide were then added. The reaction mixture was shaken in the dark for 2 days; then 10 ml of water were added and the solution was heated to 40.degree. The reaction mixture was again shaken for one hour; a further 75 ml of water were then added and the solution was freed from insoluble dicyclohexylurea formed by filtration.

  The filtrate was evaporated, diluted with 50 ml of water and evaporated again to remove the residual pyridine. The residue obtained in this way was partitioned between water and ether, 150 ml (1: 1), the aqueous portion was freed from ether in vacuo after the second extraction. The remaining aqueous solution (90 ml) was treated with 2.16 g (90 mmol) of lithium hydroxide. The solution was heated to 100 ° C. for one hour. The suspension was then cooled and lithium phosphate was removed by filtration. The solid substance was washed with 0.01N lithium hydroxide solution; the wash water was combined with the filtrate.

  The filtrate was adjusted to a pH of 7 by adding an acidic exchange resin [ <(Dowex 50 (H +)]. The mixture was then filtered again to remove the resin. The solution was evaporated to a volume of 25 ml, operating under reduced pressure at 40 C. The solution was then over 75 ml fresh Dowex 50 resins were added and the resin was eluted with water until the pH of the eluate was in the range of 4 - 5. The pH of the solution obtained was adjusted to 7.5 by adding concentrated ammonium hydroxide Solution containing product, about 200 ml, was absorbed on a column charged with Dowex AG-1 (formate) (125 ml), the column was eluted with 125 ml of water.



  The column was then eluted a second time with 0.02 M formic acid solution; this eluate was collected in 20 ml fractions at a flow rate of 2 ml per minute. 200 ml forerun were discarded: fractions 13-33 were combined and lyophilized. In this way, a white crystalline precipitate was obtained in an amount of 250 mg.



  This material was recrystallized twice from water at 40 ° C. At the end of the treatment, compound 68 was in the form of fine needles.



  Analysis for C9H14O8N5P:
Calc .: C 33.44 H 4.37 N 13.00 P 9.58
Found: C 33.37 H 4.88 N 12.61 P 9.75
Example 69
N4-Benzoyl-1-ss-D-arabinofuranosylcytosine-5'-phosphate (69)
A solution of 50 mmol of pyridinium 2-cyanethyl phosphate in 10 ml of dry pyridine was prepared. 2.77 g of N4-benzoyl-1- (2 ', 3'-di-O- -benzoyl-, 8-D-arabinofuranosyl) cytosine were added to this solution and the solution was evaporated to dryness. The mixture was dissolved in 25 ml of pyridine, treated with 3.09 (150 mmol) of dicyclohexylcarbo diimide and shaken for 51/2 days at room temperature. Then 15 ml of water were added and the mixture was extracted twice with SSB. The insoluble urea compound formed was filtered off.

  The solution was diluted with 40 ml of pyridine cooled to about 0 C with ice and adjusted to a sodium hydroxide content of about In by adding about 40 ml of ice-cold 2N sodium hydroxide solution. The reaction was terminated after 20 minutes by adding an excess of pyridinium Dowex 50 X 8.



  The resin was filtered off and washed with water; the washing waters and the filtrate were combined and, after the addition of 200 mg ammonium bicarbonate, evaporated to about 25 ml under reduced pressure. The precipitate formed in the 25 ml solution was filtered off. The filtrate was evaporated under reduced pressure: the residue was taken up in a solvent mixture consisting of 1 m ammonium acetate (pH 6) and isopropyl alcohol in a ratio of 2: 5. The solution was absorbed on a cellulose column with a column volume of 1850 ml. The column was packed with the same mixed solvent as mentioned above.



  A solvent mixture was used to elute the column, which consisted of a 1 molar aqueous ammonium acetate solution and isopropyl alcohol in a ratio of 2: 5. The first 600 ml of the eluate were discarded. Then 20 ml fractions were collected for a total of 325 fractions. Fractions 55-110 were combined and contained about 90% of the theoretically possible amount of compound 69. These fractions were concentrated to a small volume in the presence of 10 ml of pyridine. The residue was diluted with water to a volume of 50 ml; this solution was absorbed on a column charged with pyridinium Dowex 50 X 8. 3 liters of deionized water was used to elute the column.

  The entire solution which ran off was concentrated under reduced pressure, diluted four times with 1% strength aqueous pyridine solution and in each case concentrated again. Finally the residue was taken up in dilute aqueous pyridine and lyophilized twice from this solvent. In this way, compound 69 was obtained as a white solid substance in a yield of 1.81 g (70%).

 

  Analysis for C16H18N1O8P H20 pyridine:
Calc .: P 5.95
Found: P 6.06
If this solvate is heated to 100 ° C. in vacuo (15 mm Hg) for 72 hours, the pure compound 69 is obtained. In the manner described in Example 68, other N-acyl-1- (2 ', 3'-di-O -acyl-ss-D-arabino furanosyl) cytosines are phosphorylated, whereby they lose the acyl groups in the 2'- and 3'-position and the acyl group bound to the amino group of the cytosine.



   Example 70
1-ss-D-arabinofuranosylcytosine-5-phosphate (70)
If N4 - (gi - cyclopentylpropionyl) -1- [2 ', 3' -di-O- (p-cyclopentylpropionyl) - or N4-lauroyl-1- (2 ', 3) is treated in the manner described in Example 68 '-di-O-lauroyl-ss-D-arabinofuranosyl) cytosine first with 2-cyanoethyl phosphate, then with dicyclohexylcarbodiimide and finally with lithium hydroxide at reflux temperature, the compound 70 is obtained in both cases.



   Example 71
Other 1-ss-D-arabinofuranosylcytosine-5'-phosphates
The following compounds can also be used as starting material for the procedure according to Example 68:
The following abbreviations are used in the following lists: AF =, 8-D-arabinofuranosyl RF = ss-D-ribofuranosyl DRF = ss-D-deoxyribofuranosyl N4-decanoyl-1- (2 ', 3'-di-O-decanoyl- and N4-propionyl-1- (2 ', 3'-di-O-propionyl-AF) cytosine; 1- (2', 3'-di-O-butyryl-AF) uracil; N4-phenylacetyl-1- ( 2 ', 3'-di-O-phenylacetyl-, N4-hexanoyl-1- (2', 3'-di-O-hexanoyl) - and N4-phenylpropionyl-1- (2 ', 3'-di-O -phenylpropionyl-AF) cytosine; N5-anisoyl-1- (2 ', 3'-di-O-anisoyl-AF) adenine; N4-octanoyl-1- (2', 3'-di-O-octanoyl-AF ) -3-methylcytosine (N4-butyryl-1-di-O-butyryl-AF) -5-methylcytosine; Nê-anisoyl-1- (2 ', 3'-di-O-anisoyl-AF) guanine;

   9- (2 ', 3'-di-O-lauroyl-RF) xanthine; N4-octanoyl-1- (2 ', 3'-di-O-octanoyl-AF) -3-methylcytosine; 1- (2 ', 3'-di-O-decanoyl-RF) thymine; N4-butyryl-1- (2 ', 3'-di-O-butyryl-AF) -5-methylcytosine; 9- (2 ', 3'-di-O-phenylacetyl-AF) -6-mercaptopurine; N6-propionyl-1- (2 ', 3'-di-O-propionyl-AF) -5-methyl cytosine; 1- [2 ', 3'-di-O- (ss-cyclopentylpropionyl) -RF] -5-ioduracil 1- (2', 3'-di-O-anisoyl-RF) -5-fluorouracil; 1- (3'-O-valeryl-DRF) uracil; 1- (2 ', 3'-Di-O-benzoyl-RF) -5-trifluoromethyluracil or 1- (3'-O-hexanoyl-DRF) thymine.



  This reaction gives, for example, the 5'-phosphates of l-AF-adenine, -3-methylcytosine, -5-methylcytosine and -uracil; of 9-AF-xanthine, hypoxanthine, -thymine and -6-mercaptopurine; of 1-AF-5-fluorouracil, -5-chloruracil, -5-bromuracil, -5-ioduracil and -5-trifluoromethyluracil; of l-RF-5-fluorouracil and -5-trifluoromethyluracil; of 9-RF-xanthine and guanine; of 1-R uracil, cytosine and thymine; of 9-RF-adenine, -xanthine and -6-mercaptopurine; from 1-RF-5-ioduracil; 1-DR furacil and thymine; of 9-DRF-hypoxanthine, -thymine and-adenine; l-DRP-cytosine; 9-DRF-guanine; 1-DRF-5 fluorouracil; 9-DRF-xanthine et al.



   Example 72
N4-Benzoyl-1- (2 ', 3'-di-O-acetyl-ss-D-arabinofuranosyl) cytosine-5'-phosphate (72)
A solution of N4-benzoyl-1-ss-D-arabinofuranosylcytosine-5'-phosphate (that is the compound 69 prepared according to Example 69) was suspended in a mixture of 15 ml of pyridine and 15 ml of acetic anhydride. This mixture was stirred for 18 hours at room temperature. The solution was then diluted with 15 ml of water and stirred for a further 3 hours at room temperature. The solvent was then removed at 30 ° C. in a high vacuum; the residue was triturated with ether. A rubber-like material was obtained, which was freed from ether in a vacuum.

  The residue was taken up in dry pyridine; the solution was turned off at 4 ° C.



  The solid product formed during this treatment was filtered off; it was the pure compound 72.



   Example 73
N4-Benzoyl-1- (2 ', 3'-di-O-propionyl-ss-D-arabinofura nosyl) cytosine-5'-phosphate (73)
To prepare compound 73, compound 69 was reacted with propionic anhydride in pyridine in the manner described in Example 72.



     Example 74
N4-Benzoyl-1- (2 ', 3'-di-O-butyryl-ss-D-arabinofura nosyl) cytosine-5'-phosphate (74)
To prepare compound 74, compound 69 was reacted with butyric anhydride in pyridine in the manner described in Example 72.



   Example 75
N4-Benzoyl-1- (2 ', 3'-di-O-benzoyl-ss-D-arabino furanosyl) cytosine-5'-phosphate (75)
This compound is obtained by reacting compound 69 with benzoic anhydride in pyridine in the manner described in Example 72.



   Example 76 N4-Benzoyl-1-di-O-phenyacetyl-AF) cytosine -5'-phosphate (76)
This compound is obtained by reacting compound 69 with phenylacetic anhydride in pyridine in the manner described in Example 72.



   In each of the following Examples 77-105, the procedure described in Example 72 is used and pyridine is used as the solvent. Examples 77 to 105 are compiled in a table. If an * is given after the acylating agent in the table, this means that the acylating agent is used in excess over the other reaction component.



   In the table below, + = 2 ', 3'-di-O is-5'-phosphate of acylating agent such as 5'-phosphate of game +
77 N4-anisoyl-1-AF-cytosine valeric anhydride N4-anisoyl-1 (# - valeryl-AF) -cytosine
78 l-AF-cytosine acetic anhydride and N6-acetyl-1 - (# - acetyl-AF) -cytosine
Tetraethylammonium acetate *
79 l-AF-cytosine propionic anhydride * N4-propionyl-1 - (# - propionyl-AF) -cytosine
80 l-AF-adenine phenylpropionic anhy- N6-phenylpropionyl-9 - (# - phenylpropionyl-AF) - drid * -adenine
81 l-AF-uracil Benzoic anhydride * l - (+ - benzoyl-AF-uracil
82 l-AF-xanthine hexanecarboxylic anhydride * 9 - (+ - heganoyl-AF) -xanthine
83 1-AF-6-mercaptopurine ss-Cyclopentylpropionyl- <RTI

    ID = 20.8> 9 - [# (ss-cyclopentylpropionyl) -AF] -6-mercapto chloride * purine
84 9-AF-guanine lauroyl chloride * Nê-lauroyl-9 - (# - lauroyl-AF) -guanine
85 l-AF-thymine decanoyl chloride * l - (+ - decanoyl-AF) -thymine
86 N4-Benzoyl-1 -RF-cytosine propionic anhydride N4-Benzoyl-1 - (# - propionyl-RF) -cytosine
87 N4-Benzoyl-1-RF-cytosine butyric anhydride N4-Benzoyl-1 - (# - butyryl-RF) -cytosine
88 N4-Benzoyl-1-RF-cytosine Benzoic anhydride N4-Benzoyl-1 - (# - benzoyl-RF -) - cytosine
89 N6-Benzoyl-9-RF-adenine. Propionic anhydride N6-Benzoyl-9 - (# - propionyl-RF) -adenine
90 l-RF-uracil butyric anhydride 1 - (- butyryl-RF) -uracil
91 9-RF-6-mercaptopurine

   Benzoic anhydride 9 - (+ - Benzoyl-RF) -6 mercaptopurine
92 1-RF-5-fluorouracil Phenylpropionic an- 1 - (# - Phenylpropionyl-RF) -5-fluorouracil hydride
93 N2-Acetyl-1-RF-guanine Octanecarboxylic anhydride Nê-Acetyl-1 - (# - octanoyl-RF) -guanine
94 l-RF-thymine Benzoic anhydride 1 - (# - Benzoyl-RF) -thymine
95 N4-Benzoyl-1-DRF-cy-propionic anhydride N4-Benzoyl-1- (3'-O-propionyl-DRF) -cytosin tosine
96 N4-Benzoyl-1-DRF-cy-butyric anhydride N4-Benzoyl-1- (3'-O-butyryl-DRF) -cytosin tosine
97 N4-Benzoyl-1-DRF-cy-benzoic anhydride N4-Benzoyl-1- (3'-O-benzoyl-DRF) -cytosin tosine
98 N6-Benzoyl-9-DRF-ade-propionic anhydride N6-Benzoyl-9- (3 '-O-propionyl-DRF) -adenine nin
99 N2-phenylacetyl- 1 -DRF- butyric anhydride

   Nê-Phenylacetyl-1- (3'-O-butyryl-DRF) -guanine guanine 100 l-DRF-uracil Benzoic anhydride 1- (3'-O-Benzoyl-DRF) -uracil 101 9-DRF-hypoxanthine valeric anhydride 9- ( 3 '-O-Valeryl-DRF) -hypoxanthine 102 N4-Propionyl-1-DRF-3-Butyric anhydride N4-Propionyl-1- (3'-O-butyryl-DRF) -3-mesthylcymethylcytosin tosine 103 1 -DRF- 5'-trifluoromethyl-lauroyl chloride 1- (3 '-O-Lauroyl-DRF) -5'-trifluoromethyluracil uracil 104 I-DRF-thymine hexanecarboxylic anhydride 1- (3'-O-hexanoyl-DRF) -thymine 105 1-DRF- 6-mercaptopurine anisoyl chloride 9- (3'-O-anisoyl-DRF) -6-mercaptopurine
Example 106 N4-Benzoyl-1 - (TPM-AF) cytosine (106) A.

  N4-benzoyl-l-AF-cytosine (106a)
0.4 g of sodium hydroxide is added to a solution of 0.5 g of N4-benzoyl-1- (2 ', 3'-di-O -benzoyl-AF) cytosine in 17 ml of ethanol which has been cooled to 50 ° C. 3 ml of water. This mixture is left to stand at 5 ° C. for 30 minutes and then poured into 80 ml of ice water. The aqueous solution is neutralized with in hydrochloric acid and filtered. The separated precipitate is washed on the filter with water and then recrystallized twice from ethanol-SSB. Compound 106a is obtained in this way.



   B. Compound 106 is obtained by treating compound 106a with triphenylchloromethane (TPCM) for eight days at 22-250C in the manner described in Example 1.



   Example 107 N6-Butyryl-1- (TPM-AF) adenine (107) A. N6-Butyryl-1-AF-adenine (107a)
In the manner described in Example 106, NG - butyryl -1 - (2 ', 3'-di-O-butyryl-AF) cytosine is treated at 50 ° C. with sodium hydroxide. In this way connection 107a is obtained.



   B. Compound 107 can be obtained by treating compound 107a with TPCM for eight days at 250C in the manner described in Example 1.



   Example 108 N2-Benzoyl-9- (TPM-AF) gmanine (108) A. N2-Benzoyl-9-AF-guanine (108a)
In the manner described in Example 106, Ne-benzoyl-9- (2 ', 3'-di-O-benzoyl-AF) quanine (that is the compound of Example 42) is treated at 5 ° C. with sodium hydroxide solution. In this way the connection 108a is obtained.



   B. Compound 108 is obtained when compound 108a is treated with TPCM for eight days at 22-250C in the manner described in Example 1.



   Example 109
N4-valeryl-1- (5'-TMP-AF) -5-methylcytosine (109) A. N4-valeryl-l-AF-5-methylcytosine (109a)
The compound 109a is obtained if N4-valeryl-1- (2 ', 3' -di-O-valeryl-AF) -5-methylcytosine is treated with sodium hydroxide at 5 ° C. in the manner described in Example 106.



   B. Compound 109 can be obtained from compound 109a if the latter is treated with TPCM for eight days at 22-250C in the manner described in Example 1.



   Example 110
N4-Lauroyl-1- (TPM-AF) -3-methylcytosine (110) A. N4-Lauroyl-1-AF-3-methylcytosine (110 a)
N4-Lauroyl-1- (2 ', 3'-di-O-lauroyl-AF) -3-methylcytosine is treated at 50 ° C. with sodium hydroxide in the manner described in Example 106.



   B. Compound 110 can be obtained from compound 110a by treating the latter in the manner described in Example 1 for eight days at 22-250C with TPCM.



   Example III NC-Benzoyl-l- (TPM-RF) cytosine (111) A. N4-Benzoyl-l-RF-cytosine (purple)
N4-benzoyl-1- (2 ', 3'-di-O-benzoyl-RF-cytosine is treated with sodium hydroxide at 50 ° C. in the manner described in Example 106. The compound lavender is obtained in this way.



   B. The compound 111 can be obtained from the compound lilac by treating the latter in the manner described in Example 1 for eight days at 22-250C with TPCM.



   Example 112 N6-Acetyl-9- (TPM-RF) adenine (112) A. N6-Acetyl-9-RF-adenine (112a)
N6-acetyl-9- (2 ', 3'-di-O-acetyl-RF) adenine is treated with sodium hydroxide at 50 ° C. in the manner described in Example 106. In this way connection 112a is obtained.



   B. Compound 112 can be obtained from compound 112a by treating the latter in the manner described in Example 1 at 22-250C for eight days with TPCM.



   In the manner described in Example 106, other compounds of the formula VIII can also be obtained if a product of the formula VII is reacted with a base to form a compound VIIa and this product is etherified. The following compounds of the formula VIII can be obtained: N4-acetyl-, -propionyl-, -Butyryl-, -Valeryl-, -hexanoyl-, -Octanoyl-, -Lauroyl- and -phenylacetyl-1- (TPM-AF) cytosine; N4-lauroyl-1- (TPM-RF) cytosine; N6-phenyl-acetyl-9- (TPM-RF) adenine; N4-benzoyl-1 - (TPM-RF) -3-methylcytosine; N4-heptanoyl-1 - (TPM-RF) -5-methylcytosine; N2-butyryl-9- (TPM-RF) guanine; N2-anisoyl-9- (TPM-RF) guanine; N2-decanoyl-9- [5'-O- (p-methoxyphenyl) diphenylmethyl-
RF] adenine;

  ; N4-propionyl-1- [5'-O- (p-methoxyphenyl) diphenylmethyl- -RF] -5-methylcytosine; N4-phenylacetyl-1- [5'-O- (p-methoxyphenyl) diphenyl methyl-DRF] -3-methylcytosine et al.



   Example 113
N4-anisoyl-1- [5- (p-methoxyphenyl) diphenylmethyl
AF] cytosin-3'-yl-1- (3'-O-acetyl-DRF) uracil-5'-yl phosphate (XIII); N4-anisoyl-1- [5 '-) p-methoxyphenyl) diphenylmethyl-AF \ cytosine-2-yl-1- (3-0-acetyl-DRF) -uracil-5'-yl-phosphm (XIV); 1-AF-cytosine-2'-yl-1-DRF-uracil-5'-yl-phosphate (XV);
1-AF-cytosine-3'yl-1-DRF-uracil-5'-yl-phosphate (XVI)
EMI22.1
  
EMI23.1
  
EMI24.1
  
A mixture of 900 mg (1.38 mmoles) N4-anisoyl -1 [5'-p-methoxyphenyl) diphenylmethyl-AF] cytosine and 1.38 mmoles 1- (3'-acetyl-DRF) uracil-5'-yl phosphate in 2 ml of pyridine is rendered anhydrous by repeated evaporation with anhydrous pyridine under reduced pressure.

  The dry residue is taken up in 16 ml of dry pyridine, and 3 g of dicyclohexylcarbodiimide are added. The mixture is shaken in a closed container in the dark and at room temperature (22-260C) for five days. Then 6 mol of aqueous pyridine - 1 part of water to 2 parts of pyridine - are added and shaken again. The insoluble urea is filtered off; this precipitate is washed with 20 ml of pyridine. The filtrate and the washing water are extracted twice with a mixture of cyclohexane and ether in a ratio of 1: 1. The cyclohexane-ether mixture is discarded; the pyridine solution is concentrated to a small volume in vacuo at 300C.

  The residue is diluted again with 20 ml of pyridine and concentrated again to about 5 ml. Thereafter, it is chromatographed on diethylamino ethyl cellulose with 50% ethanol-50% buffer and an aqueous triethylamine acetate solution (pH 7.5), the concentration of which rose from 0.02 to 1 m. In this way, a mixture of the compound XIII and the compound XIV was obtained. The compounds XIII and XIV can be separated by countercurrent distribution according to Craig.



   To obtain the non-acylated compound, a corresponding solution (5 ml, as above) was made up to 25 ml with pyridine and treated with 60 ml of alcoholic ammonia (3 parts of concentrated ammonium hydroxide to 1 part of ethanol). The homogeneous solution thus obtained is left to stand at room temperature for 2 days; then it is concentrated to dryness at 300C in a vacuum. The residue is taken up with 25 ml of 80% acetic acid. The mixture is left to stand for 35 hours at room temperature and the acid is then removed using a high vacuum pump. The remaining residue is suspended in 20 ml of water. The suspension is adjusted to a pH of 8 with 3N ammonium hydroxide. The water-insoluble material is extracted twice with ether. The ether extracts are discarded.



  The aqueous solution (27 ml) is absorbed on a DEAH cellulose (carbonate) column with a column volume of 770 ml. The column is washed with 60 ml of water, which is discarded. A triethylamine carbonate solution (pH 7.5), the concentration of which increases from 0 to 0.25m, is used for elution.



  20 ml fractions are collected at a flow rate of 0.5 to 1.5 ml per minute. Fractions 110 to 140 are combined; they contain the desired material and unconverted l-ss-D-deoxyribofuranosyluracil. After evaporation, the desired compound can be obtained in 28% yield from these fractions. In order to separate the 1-8-D-deoxyribofuranosyluracil from the mixture, the entire product was chromatographed on fiber paper (Whatman 3MM). The paper was prewashed with a mixed solvent of 7 parts isopropyl alcohol, 1 part concentrated ammonium hydroxide, and 2 parts water. The same mixture of solvents was used for the chromatography, approximately 500 optical density units (about 25 mg) of the material was applied to each 6 "sheet.

  The dinucleoside phosphate areas (Rf 0.36, determined with the aid of UV light) were cut out and eluted with water. In this way, 4,620 optical density units of mixed l-AF-cytosin-3-yl, 1 -DRF-uracil-5-yl-phosphate and 1-AF-cytosin-2-yl, 1 -DRF-uracil- 5'-yl phosphate can be isolated in aqueous solution. This was stored in a frozen state at a pH of 4.5. A Dowex 1 X 2 (formate) column with a column volume of 50 ml, which is described in the J. Org.

  Chem. 29, 1078 (1964) was prepared with a portion of the above-described aqueous solution, which contained 2100 optical density units at 265 in, at a pH of 8.5 (adjusted with n-ammonium hydroxide ) loaded. The column was eluted with a sodium formate solution (pH 5.0), the concentration of which increased from 0.04 to 0.055 M sodium formate solution, using 1 liter of each salt solution. The column fractions were analyzed at 265 mg using a Vanguard 1056 o.n., ultraviolet monitor. 20 ml fractions were collected at a drop rate of 1.50 ml per minute. The separation of the dinucleoside phosphates was quantitative.

  Fractions 41 to 49 (330 optical density units at 265 ml) contained 1-AF-cythosin-2'-yl-1-DRF-uracil-5'-yl-phosphate XV in about 25% yield in the mixture. Compound XV (270 optical density units at 269 mF and pH 3.5) was resistant to snake venom diesterase, spleen diesterase, deoxyribonuclease and ribonuclease.

  Fractions 61 to 73 consisted of 1-AFcytosin-3'-yl-1-DRF-uracil-5'-yl-phosphate (XVI) (965 optical density units, 269 mu at pH 2.0); after evaporation a light white powder remained, which in the analysis gave the following values: Analysis for Cr8H24N5012P 4H2O:
Calc .: C 35.70 H 5.49 N 11.57
Found: C 35.70 H 5.33 N 10.62
This material (XVI) was resistant to ribonuclease and deoxyribonuclease, but was degraded to 1-AF -cytosine-3-phosphate and to l-DRF-uracil by spleen diesterase and to l-AF-cytosine by snake venom diesterase.



   Example 114
1- (TMP-DRF) thymin-3'-ylNÚ-benzoyl-1- (2 ', 3'-di-O acetyl-AF) cytosine-5' -yl-phosphate (114) and triethylamine salt of the l-DRF- thywrin-3'-yl-1-AF- -cytosin-5'-yl-phosphates (114a)
The nucleotide N4-benzoyl-1- (2 ', 3'-di-O-acetyl-AF) cytosin-5'-yl-phosphate (3.2 mmol) was evaporated to dryness several times at 40 ° C. with dry pyridine.

 

  1- (TPM-DRF) thymine (1.6 g; 3.3 mmol) and 1 g of dry pyridinium Dowex 50W X 8 (1 g) were then added to the product. The mixture was evaporated to dryness twice with dry pyridine. The organic residue obtained in this way was dissolved in 35 ml of dry pyridine, and 6.8 g of dicyclohexylcarbodiimide were added. The suspension was in the dark at room temperature, i.e. shaken at about 240C, 4 days. Then 1.5 g (3.1 mmol) of 1- (TPM-DRF) thymine was added and shaking was continued for a further 2 days. The reaction was then terminated by adding 5 ml of water. The insoluble dicyclohexylurea formed was filtered off; the precipitate was washed on the filter with 20 ml of pyridine. The pyridine solution and the washing waters were combined and evaporated in vacuo at 40.degree.

  In this way a residue was obtained which was partitioned between 200 ml of water and 100 ml of ethyl acetate. The urea was again filtered off from the suspension obtained in this way; the ethyl acetate solution was washed with water, dried over anhydrous sodium sulfate and evaporated to dryness.



  The syrupy mixture obtained in this way was taken up in 40 ml of methanol. When the methanolic solution was evaporated in vacuo, compound 114 remained as a solid substance.



   In order to obtain the non-esterified and non-etherified material, the above-mentioned phosphate was taken up in 40 ml of methanol. Concentrated ammonium hydroxide (10 ml) was added to this solution until it became cloudy. It was left to stand at room temperature for 22 hours. The solvents were then removed at 350 ° C. in a vacuum of 15 mm Hg. The residue thus obtained was suspended in 100 ml of 80% acetic acid. After 4 days the acetic acid was removed in vacuo at 350 ° C. (15 mm Hg); the residue was again suspended in 40 ml of water. The pH of this solution was adjusted to 8 with 3N ammonium hydroxide.



  The addition of the ammonium hydroxide resulted in a suspension which was extracted with ether and freed from insoluble material by filtration. The ether was removed from the remaining aqueous solution under reduced pressure. The ether-free solution was absorbed on a diethylaminoethylcellulose column which had been activated with 1.5N ammonium carbonate. The column had a capacity of 1500 ml. The column was eluted with a solution which contained amounts of triethylamine bicarbonate (pH 7) increasing from 0 to 0.12 m: 10 liters of solvent were collected in 20 ml fractions (initial column speed, 1.2 ml per minute). A forerun of 560ml was discarded.

  The fractions 130 to 330 were combined and subjected to freeze-drying: a pale colored residue was obtained in this way. This residue was redissolved in aqueous ethanol containing activated carbon (Carco C60). The solution was then filtered; The desired compound 114a could be obtained from the filtrate by freeze-drying.



  This material was redissolved and filtered through an acid exchange resin (Dowex 50W X 8). Concentrating the filtrate gave 1-DRF-thymin-3'-yl-1-AF-cytosin-5'-yl-phosphate. The substance obtained in this way was not degraded by ribonucleic acid diesterase and deoxyribonucleic acid diesterase, but completely by snake venom diesterase to l-AF-cytosine--5'-phosphate and thymidine and by spleen diesterase to l-DRF-thymine-3 ' -phosphate and l-AF-cytosine degraded.



   In the same manner as described above, the crude 1-DRF-thymin-3'-yl-1-AF-cytosin-5'-yl-phosphate can be eluted from the column with pyridine bicarbonate in the form of the pyridine salt.



   In each of Examples 115 to 128 below, the procedure explained in Example 113 was used. The dinucleoside phosphate esters were formed at room temperature in pyridine and in the presence of dicyclohexylcarbodiimide. The subsequent selective hydrolysis of the dinucleoside phosphate esters obtained in this way was carried out first with ammoniacal ethanol and then with 80% strength aqueous acetic acid.



   Example 115
First, N4-anisoyl-1- (TPM-AF) cytosine-5'-yl-phosphate is added to a mixture of dinucleoside phosphate esters, namely N4-anisoyl-1- (TPM-AF) cytosine-3 '-yl- N4-anisoyl- (2 ', 3'-di-O-benzoyl-AF) -5'-yl-phosphate and N4-anisoyl -1- (TPM-AF) cytosine -2' - yl - N4- -anisoyl- (2 ', 3' - di - O - benzoyl-AF) cytosin-5'-yl-phosphate um. The selective hydrolysis of this dinucleoside phosphate ester mixture gives 1-AF-cytosine-3'-yl-1-AF-cytosin-5'-yl-phosphate and 1 -AF-2'-y1-1 -AF-cy - tosin-5'-yl-phosphate.



   Example 116
The two dinucleoside phosphate esters N4 are first obtained from N4-benzoyl-1- (TPM-AF) ctosine and NG-decanoyl-9 - (2 ', 3' - di-O-decanoyl-RF) adenine-5'-phosphate -Benzoyl-1 - (TPM-AF) cytosin-2'-yl-N6-decanoyl-9- (2 ', 3' -di-0-decanoyl-RF) adenin-5'-yl-phosphate and N4 -benzoyl - 1 - (TPM-AF) cytosin-3'-yl-N'-decanoyl-9- (2 ', 3'- di-O-decanoyl-RF) ademin-5'-yl-phosphate, which by chromatography can be separated. The hydrolysis of the dinucleoside phosphate esters gives the free dinucleoside phosphates, namely l-AF-cytosine-2'- -yl-9-RF-adenin-5'-yl-phosphate and 1-AF-cytosin-3'-yl- -9-RF-adenin-5'-yl-phosphate.



   Example 117
The two dinucleoside phosphate esters N4 - benzoyl - are obtained from N4-benzoyl-l- (TPM-AF) cytosine and 9- (2 ', 3'-di -O-octanoyl-RF) 6-mercaptopurine-5' -phosphate 1- - (TPM - AF) cytosin 2 '- yl-9- (2', 3'-di-O-octanoyl - RF) -6 -mercaptopurin-5'-yl-phosphate and N4-benzoyl-l- ( TPM -AF) cytosin-3'-yl-9- (2 ', 3'-di-O-octanoyl-RF) -6-mercaptopurine phosphate. The ester mixture can be separated into the components by chromatography. The hydrolysis of the dinucleoside phosphate esters gives 1-AF-cytosin-2'-yl-9-RF-6-mercaptopurin-5'-yl-phosphate and 1-AF-cytosin-3'-yl-9-RF- 6-mercaptopurin-5-yl-phosphate as free dinucleoside phosphates.



   Example 118
A mixture of the two dinucleoside phosphate esters N4 -Benzoyl - is obtained from N4-Benzoyl-1- (TPM-AF) cytosine and 1- (2 ', 3'-di -O-phenylacetyl-RF) thymine-5'-phosphate 1 - (TPM-AF) cytosin-2'-yl-1- (2 ', 3'-di-O.phenyl-acetyl-RF) thymin-5'-yl-phosphate and N6-benzoyl-1 - (TPM -AF) cytosine-3'-y1-1 - (2 ', 3'-di-O-phenylacetyl-RF) -thymin-5'-yl-phosphate. These two esters can be separated by chromatography. The hydrolysis of the two dinucleoside phosphate esters gives the free dinucleoside phosphates l-AF-cytosin-2'-yl and 1 -AF-cytosin-3-yl- 1 -; p-D-ribofuranosylthymin-5'-yl-phos- phat.

  If the mixture is present, the free dinucleoside phosphates can also be separated by chromatography, as in all other cases.



   Example 119
From 1- (TMP-RF) -5-fluorouracil and N4-benzoyl-1
3'-di-O-benzoyl-AF) -5-mlsethylcytosine-5'-phosphate, a mixture of the two dinucleoside phosphate esters l- (TPM-RF) -5-fluorouracil-2'-yl- and 1 - (TPM-RF) -5-fluorouracil-3'-yl-N4-benzoyl-1- (2 ', 3'-di-O- -benzoyl-AF) -5-methylcytosine-5'-yl-phosphate. Hydrolysis of the dinucleoside phosphate esters gives the free dinucleoside phosphates, namely l-RF-5-fluorouracil-2'-yl and 1-RF-5-fluorouracil-3'-yl-AF-5-methylcy tosin-5 ' -yl phosphate.



   Example 120
9- (TPM-AF) xanthine is reacted with N2-decanoyl-9- (2 ', 3'-di-O-decanoyl-RF) quanine-5'-phosphate, so a mixture of the two dinucleoside phosphate esters 9- is obtained. (TPM-AE) xanthln-2'-yi- and 9- (TPM-AF) xanthin-3'-yl-N2- decanoyl-9- (2 ', 3'-di-O-decanoyl-RF) guanine - 5'-yl phosphate. This mixture can optionally be separated into the components by chromatography. In the hydrolysis of the dinucleoside phosphate esters, the free dinucleoside phosphates, namely 9-AF-xanthine-2'- or -3'-yl-1-RF-guanin-5'-yl-phosphate, are obtained.



   Example 121
The two dinucleoside phosphate esters N6-anisoyl-9 are first prepared from N6-anisoyl-9- (TPM-AF) adenine and 1- (3'-O -benzoyl-DRF) -5-trifluoromethyl-uracil-5'-phosphate - (TPM-AF) adenine-2 'u. -3'-yl-1- (3'-O-DRF) - -5-trifluoromethyluracil-5'-yl-phosphate. The hydrolysis of these two dinucleoside phosphate esters gives a mixture of 9-AF-adenine-2'- or -3'-yl-1-DRF-5-trifluoromethyluracil-5'-yl phosphate.



   Example 122
The dinucleoside phosphate ester N4 is obtained from N4-anisoyl-1- (TPM-DRF) cytosine and N4-anisoyl-1- (2 ', 3'-di-O-benzoyl-RF) cytosin-5'-yl phosphate -Anisoyl-1- - (TPM-DRF) cytosine - 3'-yl - N4 - anisoyl-1- (2 ', 3'-di-O-benzoyl-RF) cytosin-5-yl-phosphate. In the selective hydrolysis, the dinucleoside phosphate obtained is l-DRF-cytosin-3-yl-1-RF-cytosin-5-yl-phosphate.



   Example 123
From N6-anisoyl-9- [5'-O- (p-methoxyphenyl) diphenylmethyl-DRF] adenine and N4-benzoyl-1- (2 ', 3'-di-O-benzoyl-RF) -3-methyl- cytosine 5'-phosphate is used as the dinucleoside phosphate ester N6-anisoyl- 9- [3'-O- (p-meRh- oxyphenyl) diphenylmethyl-DRF) adenin-3'-yl-N4-benzoyl- (2 ' , 3'-di-O-benzoyl-RF) -3-methylcytosin-5-yl-phosphate.



  9 -DRF-adenin-3'-yl-1-RF-3-methylcytosin-5'-yl phosphate is obtained from these by selective hydrolysis.



   Example 124
From N4-Lauroyl-1- (TPM-DRF) -5-methylcytosine and
1- (2 ', 3'-Di-O-hexanoyl-AF) uracil-5-yl-phosphate is obtained as the dinucleoside phosphate ester N4-lauroyl-1- (TPM -DRF) -5-methylcytosin-3'-yl - 1 - (2 ', 3'-di-O-hexanoyl -AF) uracil-5'-yl-phosphate. On selective hydrolysis, this yields 1-DRF-5-methylcytosin-3'-yl-1-AF-uracil-5'-yl-phosphate.



   Example 125
From 1- [5 '-0- bis (p - methoxyphenyl) phenylmethyl -DRF] thymine and 9- (2'; 3 '-Di-O-benzoyl-AF) -6-mercaptopurin-5'-yl- phosphate is obtained first of all the dinucleoside phosphate ester 1- [5'-O-bis (p-methoxyphenyl) phenylmethyl-DRF] thymin-3'-yl-9 - (2 ', 3'-di - O-benzoyl- AF) -6-mercaptopurine-5'-y1-phosphate. On selective hydrolysis, this yields 1-DRF-thymin-3'-yl-9-AF-6-mercaptopurin-5'-yl-phosphate.



   Example 126
From 1 - (TPM-DRF) -5-ioduracil and N2-propionyl-9 - (2 ', 3'-di-O-propionyl-RF) guanin-5'-yl-phosphate, 1- (TPM- DRF) -5-ioduracil-3'-yl-Ne-propionyl-9- (2 ', 3'-di-O-propionyl-RF) guanin-5'-yl phosphate as a dinucleoside phosphate ester. On selective hydrolysis, this gives l-DRF-5-ioduracil-3'-yl-9-RF-guanine-5'-yl-phosphate.



   Example 127
By reacting l- (TPM-AF) cytosine with NG benzoyl-9- (3'-O-benzoyl-DRF) adenine-5'-phosphate, a mixture of the two dinucleoside phosphate esters l- (TPM-11F) is obtained. cytosin-2'-yl- and 1- (TPM -AF) cytosin-3'-yl-N5-benzoyl-9- (3'-O-benzoyl-DRF) adenin-5'-yl-phosphate, which may optionally be carried out by Chromatograph into the components can be separated.



  The hydrolysis of the dinucleoside phosphate esters gives 1 AF-cytosin-2-yl- and 1-AF-cytosin-3'-yl-9-DRF-adenin-5'-yl phosphate as free dinucleoside phosphates,
Example 128
By reacting l- (TPM-AF) cytosine with N2 -benzoyl-9- (2 ', 3' - di -0- benzoyl RF) guanine-5'-phosphate, you first obtain a mixture of the two dinucleoside phosphate esters l- fIPM-AF) cytosin-2'-yl- and 1- (TPM-AF) cytosin-3 '- yl - N2 - benzoyl-9- (2', 3'-di-O-benzoyl-RF) guanine-5 '-yl phosphate. On hydrolysis, these give l-AF-cytosin-2'-yl- and 1-AF-cytosin-3'-yl-9-ss-D-ribofuranosylguanin-5'-yl-phosphate as free phosphates.



   In the manner described in Example 113, other dinucleoside phosphates of the formulas XI and XII can be prepared by reacting nucleosides of the formulas II or VIII with nucleotides of the formula VI and the resulting ether esters of the formulas IX and X first with a base and then hydrolyzed with an acid. For example, the following compounds of the formulas XI and XII can be obtained: 1-AF-cytosine-5'- or. 2'-yl-1-AF-thymin-5'-yl-phosphate; 9-AF-adenine-3 'or



   -2'-yl-9-AF-xanthin-5'-yl-phosphate; 9-AF-hypoxanthin-3 'or



   -2'-yl-9-AF-mercaptoppurin-5'-yl-phosphate; l-AF-5-fluorouracil-3'- or



   -2'-yl-1-AF-5-fluorouracil-5'-yl-phosphate; l-AF-thymine-3 'or



   -2'-yl-9-RF-guanin-5'-yl-phosphate; 1 -AF-3-methylcytosine-3 'or

 

   -2'-yl-1-RF-5-chlruracil-5'-yl-phosphate; l-AF-5-ioduracil-3 'or



   -2'-yl-1 -RF-3-uracil-5'-yl-phosphate; 1 -AF-5-bromuracil-3'- or



      -2'-yl-1 -RF-3-methylcytosine-5'-yl-phosphate; l-RF-cytosine-2 'or



   -3'-yl-9-AF-xanthin-5'-yl-phosphate; l-RF-uracil-2 'or



   -3'-yl-9-AF-hydroxanthin-5'-yl-phosphate; 9-RF-6-mercaptopurine-2 'or



   -3'-yl1-AF-5-fluorouracil-5'-yl-phosphate; 9-RF-adenine-2 'or



   -3'-yl-1-AF-5-ioduracil-5'-yl-phosphate; 9-DRF-6-mercaptopurine-3'-yl-1 -AF-5-bromouracil-5'-yl-phosphate; 1-DRF4hymin-3'-yl-1 -AF-thymin-5'-yl-phosphate; 9-DRF-quanin-3'-yl-1-AF-uracil-5'-yl-phosphate; 9-DRF-trifluoromethyluracil-3'-yl-9-AF-xanthin-5'-yl-phosphate;
1 -DRF-3-methylcytosine-3'-yl-1 -AF-thymine-5'-yl-phosphate;
1-RF-5-methylcytosin-3'-yl-9-AF-adenin-5'-yl-phosphate; 1 -DRF-5-fluorouracil-3 '-yl-1 -AF - chloruracilj' -yl- -phosphate;
1-AF-5-fluorouracil-2'-yl-1-DRF-5-chloruracil-5'-yl phosphate; 1 -AF-5-fluorouracil.3'-yl-1 -DRF-5-chloruracil-5'-yl- phosphate;

   9-AF-adenine-2-yl-9-DRF-hypoxanthin-5'-yl-phosphate; 9-AF-adenin-3'-yl-9-DRF-hypoxanthin-5'-yl-phosphate; 1-AF-3-methylcytosin-2'-yl-1-DRF-5-fluorouracil-5'-yl-phosphate; 1 -AF-3-methylcytosine-3 '-yl-1 -DRF-5-fluorouracil-5' -yl- phosphate; 9-AF-xanthin-2'-yl-1-DRF-3-uracil-5'-yl-phosphate; 9-AF-xanthin-3'-yl-I-DRF-5-uracil-5'-yl-phosphate; 1-RF-3-methylcytosi-2'-yl-9-DRF-adenin-5'-yl phosphate; 1-RF-3-methylcytosin-3'-yl-9-DRF-adenin-5'-yl phosphate;
1 -RF-guanin-2'-yl-9-DRF-hypoxanthine-5'-yl-phosphate;
9-RF-guanin-3'-yl-9-DRF-hypoxanthin-5'-yl-phosphate; l-RF-5-bromuracil-2 'or

 

   -3'-yl-1-DRF-5-methylcytosin-5'-yl-phosphate;
9-RF-6-mercoptopurin-2'-yl-1-DRF-5-bromuracil-5 '.



   -yl-phosphate;
9-RF-6-mercaptopurin-3'-yl-1-DRF-5-bromouracil-5-yl-phosphate; 9-DRF-6-mercaptopurin-3'-yl-DRF-5-ioduracil-5'-yl-phosphate; 9-DRF-hypoxanthin-3'-yl-9-DRF-hypoxanthin-5'-yl phosphate; 3-DRF-3-uracil-3'-yl-1-DRF-thymin-5'-yl-phosphate; 1 -DRF-thymin-3'-yl-9-DRF-xanthine-5'-yl-phosphate; 9-DRF-adenin-3'-yl-1-DRF-5-ioduracil-5'-yl-phosphate; 9-DRF-hypoxanthin-3'-yl-9-DRF-hypoxanthin-5'-yl-phosphate et al.

 

Claims (1)

PATENT CLAIM Process for the preparation of 2 ', 5'- and 3', 5'-dinucleoside phosphates of the formulas EMI28.1 in which X1 denotes hydrogen, α-OH or p-OH, X2 and X3 each in the combinations H, -OH; H, ss-OH; ? -OH, ss-OH,? ; -OH or s-OH, ss-OH are present, and Y1 and Y2 are cytosin-1-yl, uracyl-1-yl, thymin-1-yl, adenin-9-yl, guanin-9-yl, 6-mercaptopurine -9-yl, uracil 3-yl, 5-fluorouracil-l-yl, 5-chloruracil-l-yl, 5-bromo uracil-l-yl, 5-iodouracil-l-yl, 5-trifluoromethyluracil l-yl, hypoxanthine-9-yl, Xanthin-9-yl, 5-methylcytosin-1-yl or 3-methylcytosin-1-yl represent, and pharmaceutically acceptable salts of these compounds, characterized in that a nucleoside phosphate of the formula EMI29.1 in which X 'denotes hydrogen, o-O-acyl or, 8-O-acyl, the acyl groups containing 2 to 12 carbon atoms and Ac' being an acyl radical having 2 to 12 carbon atoms, with a nucleoside of the formula EMI29.2 where T is triphenylethyl, (p-Methoxyphenyl) diphenylmethyl or bis (p-methoxyphenyl) phenylmethyl is, X is hydrogen, cc-OH or j3-OH and Y 'corresponds to one of the radicals mentioned for Y1 and Y2 in which amino or imino groups are acylated, in Presence of a dialkylcarbodiimide in the 3'-position, if X is hydrogen, or in the 2'- or 3'-position, if X is sc-OH or p-OH, but with the proviso that the nucleoside and the nucleoside phosphate, which are to be reacted with one another, are used in the combinations arabinofuranoside-arabinofuranoside, ribofuranoside-arabinofuranoside, deoxyribofuranoside-arabinofuranoside or ribofuranoside-deoxyribofuranoside, so that a dinucleoside of the formula EMI29.3 or. their mixtures are obtained in which X'1 and X'3 have the same meaning as X ', and that the intermediate products obtained are then hydrolyzed first with a weak base and then with an aqueous acid under mild conditions. SUBCLAIMS 1. The method according to claim, characterized in that the dialkylcarbodiimide is dicyclohexylcarbodiimide. 2. The method according to claim, characterized in that the dialkylcarbodiimide is dicyclohexylcarbodiimide and that the hydrolyzing agent used is ammonium hydroxide in a lower alcohol. 3. The method according to claim, characterized in that the compounds of formulas XI and XII are separated from one another by chromatography. 4. The method according to claim, characterized in that from N4-anisoyl-1- [5 '- (p-methoxyphenyl) diphenylmethyl-ss-D-arabinofuranosyl] cytosine and 1 - (3' - acetyl - D - deoxyribofuranosyl ) uracil-5'-yl-phosphate 1 -ss-D-arabinofuranosylcytosin-2'-yl-1 -, 8-D-deoxy-ribofuranosyluracyl-5'-yl-phosphate and lpD-arabinofuranosylcytosine-3 ' -yl-1-ss-D-deoxyriborufanosyluracil -5'-yl-phosphate. 5. The method according to claim, characterized in that from N4-benzoyl-1- (2 ', 3'-di-O-acetyl-ss-D-arabinofuranosyl) cytosin-5'-yl-phosphate and 1- (5 'Triphenylmethyl-ss-D-deoxyribofuranosyl) thymine 1-ss -D-deoxyribofuranosylthymin -3, - yl- 1 -D-arabinofuranosylcytosin-5'-yl-phosphate. 6. The method according to claim, characterized in that 1-ss-D-arabinofuranosylcytosin-3'-yl 1-ss-D-arabinofuranosylcytosin-5'-yl-phosphate and 1-ss -D-arabinofuranosylcytosin-2'-yl -1-ss-D-arabinofuranosylcytosin-5'-yl-phosphate 7. The method according to claim, characterized in that from N4-benzoyl-l- (5'-triphenylmethyl pD-arnbinofuranosyl) cytosine and N6-decanoyl-9- (2 ', 3'- di-O-decanoyl-, 3-D-ribofuranosyl) adenine-5'-phosphate 1-arabinofuranosylcytosin-2'-yl-9-ss-D-ribofuranosyl adenin-5'-yl-phosphate and 1 -pD-alabinofuranosylcytosin-3'-yl - 9 - pD-ribofuranosyiadenin-5'-yl-phosphate. 8. The method according to claim, characterized in that from N4-benzoyl-l- (5'-triphenylmethyl- -ss-D-arabinofuranosyl) cytosine and 9- (2 ', 3'-di-O-octanoyl-ss- D-ribofuranosyl) -6-mercaptopurine-5'-phosphate 1-ss -D-arabinofuranosylcytosin-2'-yl-9-ss-D-ribofuranosyl-6-mercaptopurine-5'-phosphate and 1-ss-D-arabinofurano sylcytosin -3'-yl-99-D-n-bofuranosyl-6-mercaptopurine-5'- phosphate. 9. The method according to claim, characterized in that from N4-benzoyl-l- (5'-triphenylmethyl- -ss-D-arabinofuranosylcytosine and 1- (2 ', 3'-di-O-phenylacetyl-pD-ribofuranosyl) thymine-5'-phosphate lss-D-arabinofuranosylcytosin-2'-yl-1-ss-D-ribofuranosylthymine-5'-yl-phosphate and 1-ss-D-arabinofuranosylcytosin-3'-yl-1 ss- D-ribofuranosylthymin-5'-yl-phosphate. 10. The method according to claim, characterized in that the compounds of the formula XI or XII obtained are converted into the corresponding pharmaceutically acceptable salts. 11. The method according to claim, characterized in that, if X2 is z-OH or ss-OH, a mixture of compounds of the formulas XI and XII is obtained.